Compositions and methods for intraocular delivery of fibronectin scaffold domain proteins

Abstract

The present disclosure relates to novel sustained-release intraocular drug delivery systems and improvements in the treatment of retinopathies. In particular, fibronectin scaffold domain proteins that selectively inhibit VEGFR-2 are contemplated.

Description

RELATED APPLICATIONS

[0001] This application is a continuation-in-part of U.S. patent application Ser. No. 11/448,171, entitled "Inhibitors of Type 2 Vascular Endothelial Growth Factor Receptors," filed Jun. 5, 2006, which is a continuation of International Application PCT/US04/40885, entitled "Inhibitors of Type 2 Vascular Endothelial Growth Factor Receptors," filed Dec. 6, 2004 and designating the U.S., which claims the benefit of U.S. Provisional Application No. 60/527,886, entitled "Inhibitors of Vascular Endothelial Growth Factor Receptors," filed Dec. 5, 2003. All of the teachings of the above-referenced applications are incorporated herein by reference.

BACKGROUND OF THE INVENTION

[0002] The present disclosure relates to novel sustained-release intraocular drug delivery systems and methods for using these systems to inhibit biological activities in the eye. In particular, the systems of the invention inhibit biological activities mediated by vascular endothelial growth factors (VEGFs).

[0003] Angiogenesis is the process by which new blood vessels are formed from pre-existing capillaries or post capillary venules; it is an important component of many physiological processes including ovulation, embryonic development, wound repair, and collateral vascular generation in the myocardium. Angiogenesis is also central to a number of pathological conditions such as tumor growth and metastasis, diabetic retinopathy, and macular degeneration. In many instances, the process begins with the activation of existing vascular endothelial cells in response to a variety of cytokines and growth factors. In cancer, tumor released cytokines or angiogenic factors stimulate vascular endothelial cells by interacting with specific cell surface receptors. The activated endothelial cells secrete enzymes that degrade the basement membrane of the vessels, allowing invasion of the endothelial cells into the tumor tissue. Once situated, the endothelial cells differentiate to form new vessel offshoots of pre-existing vessels. The new blood vessels provide nutrients to the tumor, facilitating further growth, and also provide a route for metastasis.

[0004] To date, numerous angiogenic factors have been identified, including the particularly potent factor VEGF. VEGF was initially purified from the conditioned media of folliculostellate cells and from a variety of cell lines. More recently a number of structural homologs and alternatively spliced forms of VEGF have been identified. The various forms of VEGF bind as high affinity ligands to a suite of VEGF receptors (VEGFRs). VEGFRs are tyrosine kinase receptors, many of which are important regulators of angiogenesis. The VEGFR family includes 3 major subtypes: VEGFR-1, VEGFR-2 (also known as Kinase Insert Domain Receptor, "KDR", in humans), and VEGFR-3. Among VEGF forms, VEGF-A, VEGF-C and VEGF-D are known to bind and activate VEGFR-2.

[0005] VEGF, acting through its cognate receptors, can function as an endothelial specific mitogen during angiogenesis. In addition, there is substantial evidence that VEGF and VEGFRs are up-regulated in conditions characterized by inappropriate angiogenesis, such as cancer. As a result, a great deal of research has focused on the identification of therapeutics that target and inhibit VEGF or VEGFR.

[0006] Vascular diseases of the eye comprise a major cause of blindness and have only imperfect methods of treatment. These diseases include various retinopathies and macular degeneration. Retinopathy frequently results in blindness or severely limited vision due to unorganized growth and/or damage to retinal blood vessels. There are two major types of retinopathy: diabetic retinopathy and retinopathy of prematurity. Diabetic retinopathy affects nearly 80% of all diabetics who have had diabetes for more than 15 years. Retinopathy of prematurity is thought to result from oxygen toxicity, with about 15,000 premature infants a year being diagnosed with ROP in the United States alone. Macular degeneration results from the neovascular growth of the choroid vessel underneath the macula. There are two types of macular degeneration: dry and wet. While wet macular degeneration only comprises 15% of all macular degeneration, nearly all wet macular degeneration leads to blindness. In addition, wet macular degeneration nearly always results from dry macular degeneration. Once one eye is affected by wet macular degeneration, the condition almost always affects the other eye.

[0007] Current therapeutic approaches that target or inhibit VEGF or VEGFR include antibodies, peptides, and small molecule kinase inhibitors. Of these, antibodies are the most widely used for in vivo recognition and inhibition of ligands and cellular receptors. Highly specific antibodies have been used to block receptor-ligand interaction, thereby neutralizing the biological activity of the components, and also to specifically deliver toxic agents to cells expressing the cognate receptor on its surface. Although effective, antibodies are large, complex molecules that rely on expression in recombinant mammalian cells for production. Antibodies also cause a variety of side effects that are often undesirable, including activation of complement pathways and antibody-directed cellular cytotoxicity. As a result, there remains a need for effective therapeutics that can specifically inhibit VEGF/VEGFR pathways as a treatment for disorders characterized by inappropriate angiogenesis, in particular for the treatment of retinopathies. Additionally, long-lasting treatments are in need for intraocular treatments.

SUMMARY OF THE INVENTION

[0008] The application provides sustained-release intraocular drug delivery systems comprising: a therapeutic component comprising an antiangiogenic polypeptide component; and a polymeric component associated with the therapeutic component to permit the therapeutic component to be released into the interior of an eye of an individual at a therapeutically effective dosage for a period of time after the drug delivery system is placed in the eye. The therapeutic and polymeric components may be combined in an implant device or as a plurality of particles.

[0009] In some embodiments, the antiangiogenic polypeptide component comprises an antibody, antibody fragment, or an artificial antibody, such as a scaffold region based upon a fibronectin, as well as the humanized versions thereof. An artificial antibody may comprise fibronectin based "addressable" therapeutic binding molecules ("FATBIM"), such as CT322, C7S100 and C7C100. In some embodiments, the therapeutic component is selected from the group consisting of C7S100 and C7C100; and a polymeric component associated with the therapeutic component to permit the therapeutic component to be released into the interior of an eye of an individual at a therapeutically effective dosage for a period of time after the drug delivery system is placed in the eye.

[0010] In some embodiments, the antiangiogenic polypeptide component comprises a sequence selected from SEQ ID NOs: 6-183, 186-197, 199 and 241-310. In exemplary embodiments the sequence is selected from SEQ ID NO: 194 or 195. In some embodiments the polypeptide component comprises PEG.

[0011] The application further provides novel methods of treatment. In one aspect, a method of treating a retinopathy is provided, the method comprising administering, to a patient in need thereof, a therapeutically effective amount of a polypeptide that binds to human VEGFR-2, the polypeptide comprising between about 80 and about 150 amino acids that have a structural organization comprising: i) at least five to seven beta strands or beta-like strands distributed among at least two beta sheets, and ii) at least one loop portion connecting two strands that are beta strands or beta-like strands, which loop portion participates in binding to VEGFR-2, wherein the polypeptide binds to an extracellular domain of the human VEGFR-2 protein with a dissociation constant (K.sub.D) of less than 1.times.10.sup.-6 M and inhibits VEGFR-2 mediated angiogenesis. The methods of treatment also provide for administering to a patient in need thereof the sustained-release intraocular drug delivery systems of the invention.

[0012] The antiangiogenic polypeptide components may comprise single domain polypeptides. A single domain polypeptide described herein will generally be a polypeptide that binds to a target, such as VEGFR-2, and where target binding activity situated within a single structural domain, as differentiated from, for example, antibodies and single chain antibodies, where antigen binding activity is generally contributed by both a heavy chain variable domain and a light chain variable domain. The disclosure also provides larger proteins that may comprise single domain polypeptides that bind to target. For example, a plurality of single domain polypeptides may be connected to create a composite molecule with increased avidity. Likewise, a single domain polypeptide may be attached (e.g., as a fusion protein) to any number of other polypeptides. In certain aspects a single domain polypeptide may comprise at least five to seven beta or beta-like strands distributed among at least two beta sheets, as exemplified by immunoglobulin and immunoglobulin-like domains. A beta-like strand is a string of amino acids that participates in the stabilization of a single domain polypeptide but does not necessarily adopt a beta strand conformation. Whether a beta-like strand participates in the stabilization of the protein may be assessed by deleting the string or altering the sequence of the string and analyzing whether protein stability is diminished. Stability may be assessed by, for example, thermal denaturation and renaturation studies. Preferably, a single domain polypeptide will include no more than two beta-like strands. A beta-like strand will not usually adopt an alpha-helical conformation but may adopt a random coil structure. In the context of an immunoglobulin domain or an immunoglobulin-like domain, a beta-like strand will most often occur at the position in the structure that would otherwise be occupied by the most N-terminal beta strand or the most C-terminal beta strand. An amino acid string which, if situated in the interior of a protein sequence would normally form a beta strand, may, when situated at a position closer to an N- or C-terminus, adopt a conformation that is not clearly a beta strand and is referred to herein as a beta-like strand.

[0013] In certain embodiments, the disclosure provides single domain polypeptides that bind to VEGFR-2. Preferably the single domain polypeptides bind to human KDR, mouse Flk-1, or both. A single domain polypeptide may comprise between about 80 and about 150 amino acids that have a structural organization comprising: at least seven beta strands or beta-like strands distributed between at least two beta sheets, and at least one loop portion connecting two beta strands or beta-like strands, which loop portion participates in binding to VEGFR-2. In other words a loop portion may link two beta strands, two beta-like strands or one beta strand and one beta-like strand. Typically, one or more of the loop portions will participate in VEGFR-2 binding, although it is possible that one or more of the beta or beta-like strand portions will also participate in VEGFR-2 binding, particularly those beta or beta-like strand portions that are situated closest to the loop portions. A single domain polypeptide may comprise a structural unit that is an immunoglobulin domain or an immunoglobulin-like domain. A single domain polypeptide may bind to any part of VEGFR-2, although polypeptides that bind to an extracellular domain of a VEGFR-2 are preferred. Binding may be assessed in terms of equilibrium constants (e.g., dissociation, K.sub.D) and in terms of kinetic constants (e.g., on rate constant, k.sub.on and off rate constant, k.sub.off). A single domain polypeptide will typically be selected to bind to VEGFR-2 with a K.sub.D of less than 10.sup.-6M, or less than 10.sup.-7M, 5.times.10.sup.-8M, 10.sup.-8M or less than 10.sup.-9M. VEGFR-2 binding polypeptides may compete for binding with one, two or more members of the VEGF family, particularly VEGF-A, VEGF-C and VEGF-D and may inhibit one or more VEGFR-2-mediated biological events, such as proliferation of endothelial cells, permeabilization of blood vessels and increased motility in endothelial cells. VEGFR-2 binding polypeptides may be used for therapeutic purposes as well as for any purpose involving the detection or binding of VEGFR-2. Polypeptides for therapeutic use will generally have a K.sub.D of less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M, although higher K.sub.D values may be tolerated where the k.sub.off is sufficiently low or the k.sub.on is sufficiently high. In certain embodiments, a single domain polypeptide that binds to VEGFR-2 will comprise a consensus VEGFR-2 binding sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4. Preferably, such sequence will be situated in a loop, particularly the FG loop.

[0014] In certain embodiments, the single domain polypeptide comprises an immunoglobulin (Ig) variable domain. The Ig variable domain may, for example, be selected from the group consisting of: a human V.sub.L domain, a human V.sub.H domain and a camelid V.sub.HH domain. One, two, three or more loops of the Ig variable domain may participate in binding to VEGFR-2, and typically any of the loops known as CDR1, CDR2 or CDR3 will participate in VEGFR-2 binding.

[0015] In certain embodiments, the single domain polypeptide comprises an immunoglobulin-like domain. One, two, three or more loops of the immunoglobulin-like domain may participate in binding to VEGFR-2. A preferred immunoglobulin-like domain is a fibronectin type III (Fn3) domain. Such domain may comprise, in order from N-terminus to C-terminus, a beta or beta-like strand, A; a loop, AB; a beta strand, B; a loop, BC; a beta strand C; a loop CD; a beta strand D; a loop DE; a beta strand F; a loop FG; and a beta or beta-like strand G. See FIG. 22 for an example of the structural organization. Optionally, any or all of loops AB, BC, CD, DE, EF and FG may participate in VEGFR-2 binding, although preferred loops are BC, DE and FG. A preferred Fn3 domain is an Fn3 domain derived from human fibronectin, particularly the 10.sup.th Fn3 domain of fibronectin, referred to as .sup.10Fn3. It should be noted that none of VEGFR-2 binding polypeptides disclosed herein have an amino acid sequence that is identical to native .sup.10Fn3; the sequence has been modified to obtain VEGFR-2 binding proteins, but proteins having the basic structural features of .sup.10Fn3, and particularly those retaining recognizable sequence homology to the native .sup.10Fn3 are nonetheless referred to herein as ".sup.10Fn3 polypeptides". This nomenclature is similar to that found in the antibody field where, for example, a recombinant antibody V.sub.L domain generated against a particular target protein may not be identical to any naturally occurring V.sub.L domain but nonetheless the protein is recognizably a V.sub.L protein. A .sup.10Fn3 polypeptide may be at least 60%, 65%, 70%, 75%, 80%, 85%, or 90% identical to the human .sup.10Fn3 domain, shown in SEQ ID NO:5. Much of the variability will generally occur in one or more of the loops. Each of the beta or beta-like strands of a .sup.10Fn3 polypeptide may consist essentially of an amino acid sequence that is at least 80%, 85%, 90%, 95% or 100% identical to the sequence of a corresponding beta or beta-like strand of SEQ ID NO: 5, provided that such variation does not disrupt the stability of the polypeptide in physiological conditions. A .sup.10Fn3 polypeptide may have a sequence in each of the loops AB, CD, and EF that consists essentially of an amino acid sequence that is at least 80%, 85%, 90%, 95% or 100% identical to the sequence of a corresponding loop of SEQ ID NO:5. In many instances, any or all of loops BC, DE, and FG will be poorly conserved relative to SEQ ID NO:5. For example, all of loops BC, DE, and FG may be less than 20%, 10%, or 0% identical to their corresponding loops in SEQ ID NO:5.

[0016] In certain embodiments, the disclosure provides a non-antibody polypeptide comprising a domain having an immunoglobulin-like fold that binds to VEGFR-2. The non-antibody polypeptide may have a molecular weight of less than 20 kDa, or less than 15 kDa and will generally be derived (by, for example, alteration of the amino acid sequence) from a reference, or "scaffold", protein, such as an Fn3 scaffold. The non-antibody polypeptide may bind VEGFR-2 with a K.sub.D less than 10.sup.-6M, or less than 10.sup.-7M, less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M. The unaltered reference protein either will not meaningfully bind to VEGFR-2 or will bind with a K.sub.D of greater than 10.sup.-6M. The non-antibody polypeptide may inhibit VEGF signaling, particularly where the non-antibody polypeptide has a K.sub.D of less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M, although higher K.sub.D values may be tolerated where the k.sub.off is sufficiently low (e.g., less than 5.times.10.sup.-4 s.sup.-1). The immunoglobulin-like fold may be a .sup.10Fn3 polypeptide.

[0017] In certain embodiments, the disclosure provides a polypeptide comprising a single domain having an immunoglobulin fold that binds to VEGFR-2. The polypeptide may have a molecular weight of less than 20 kDa, or less than 15 kDa and will generally be derived (by, for example, alteration of the amino acid sequence) from a variable domain of an immunoglobulin. The polypeptide may bind VEGFR-2 with a K.sub.D less than 10.sup.-6M, or less than 10.sup.-7M, less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M. The polypeptide may inhibit VEGF signaling, particularly where the polypeptide has a K.sub.D of less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M, although higher K.sub.D values may be tolerated where the k.sub.off is sufficiently low or where the k.sub.on is sufficiently high. In certain preferred embodiments, a single domain polypeptide having an immunoglobulin fold derived from an immunoglobulin light chain variable domain and capable of binding to VEGFR-2 may comprise an amino acid sequence selected from the group consisting of: SEQ ID NOs:241-310.

[0018] In certain preferred embodiments, the disclosure provides VEGFR-2 binding polypeptides comprising the amino acid sequence of any of SEQ ID NOs:192-194. In the case of a polypeptide comprising the amino acid sequence of SEQ ID NO:194, a PEG moiety or other moiety of interest, may be covalently bound to the cysteine at position 93. The PEG moiety may also be covalently bonded to an amine moiety in the polypeptide. The amine moiety may be, for example, a primary amine found at the N-terminus of a polypeptide or an amine group present in an amino acid, such as lysine or arginine. In certain embodiments, the PEG moiety is attached at a position on the polypeptide selected from the group consisting of: a) the N-terminus; b) between the N-terminus and the most N-terminal beta strand or beta-like strand; c) a loop positioned on a face of the polypeptide opposite the target-binding site; d) between the C-terminus and the most C-terminal beta strand or beta-like strand; and e) at the C-terminus.

[0019] In certain aspects, the disclosure provides short peptide sequences that mediate VEGFR-2 binding. Such sequences may mediate VEGFR-2 binding in an isolated form or when inserted into a particular protein structure, such as an immunoglobulin or immunoglobulin-like domain. Examples of such sequences include those disclosed as SEQ ID NOs:1-4 and other sequences that are at least 85%, 90%, or 95% identical to SEQ ID NOs:1-4 and retain VEGFR-2 binding activity. Accordingly, the disclosure provides substantially pure polypeptides comprising an amino acid sequence that is at least 85% identical to the sequence of any of SEQ ID NOs:1-4, wherein said polypeptide binds to a VEGFR-2 and competes with a VEGF species for binding to VEGFR-2. Examples of such polypeptides include a polypeptide comprising an amino acid sequence that is at least 80%, 85%, 90%, 95% or 100% identical to an amino acid sequence at least 85% identical to the sequence of any of SEQ ID NOs:6-183, 186-197, 199 and 311-528. Preferably such polypeptide will inhibit a biological activity of VEGF and may bind to VEGFR-2 with a K.sub.D less than 10.sup.-6M, or less than 10.sup.-7M, less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M.

[0020] In certain embodiments, any of the VEGFR-2 binding polypeptides described herein may be bound to one or more additional moieties, including, for example, a moiety that also binds to VEGFR-2 (e.g., a second identical or different VEGFR-2 binding polypeptide), a moiety that binds to a different target (e.g., to create a dual-specificity binding agent), a labeling moiety, a moiety that facilitates protein purification or a moiety that provides improved pharmacokinetics. Improved pharmacokinetics may be assessed according to the perceived therapeutic need. Often it is desirable to increase bioavailability and/or increase the time between doses, possibly by increasing the time that a protein remains available in the serum after dosing. In some instances, it is desirable to improve the continuity of the serum concentration of the protein over time (e.g., decrease the difference in serum concentration of the protein shortly after administration and shortly before the next administration). Moieties that tend to slow clearance of a protein from the blood include polyethylene glycol, sugars (e.g. sialic acid), and well-tolerated protein moieties (e.g., Fc fragment or serum albumin). The single domain polypeptide may be attached to a moiety that reduces the clearance rate of the polypeptide in a mammal (e.g., mouse, rat, or human) by greater than three-fold relative to the unmodified polypeptide. Other measures of improved pharmacokinetics may include serum half-life, which is often divided into an alpha phase and a beta phase. Either or both phases may be improved significantly by addition of an appropriate moiety. Where polyethylene glycol is employed, one or more PEG molecules may be attached at different positions in the protein, and such attachment may be achieved by reaction with amines, thiols or other suitable reactive groups. Pegylation may be achieved by site-directed pegylation, wherein a suitable reactive group is introduced into the protein to create a site where pegylation preferentially occurs. In a preferred embodiment, the protein is modified so as to have a cysteine residue at a desired position, permitting site directed pegylation on the cysteine. PEG may vary widely in molecular weight and may be branched or linear. Notably, the present disclosure establishes that pegylation is compatible with target binding activity of .sup.10Fn3 polypeptides and, further, that pegylation does improve the pharmacokinetics of such polypeptides. Accordingly, in one embodiment, the disclosure provides pegylated forms of .sup.10Fn3 polypeptides, regardless of the target that can be bound by such polypeptides.

[0021] In certain embodiments, the disclosure provides a formulation comprising any of the VEGFR-2 binding polypeptides disclosed herein. A formulation may be a therapeutic formulation comprising a VEGFR-2 binding polypeptide and a pharmaceutically acceptable carrier. A formulation may also be a combination formulation, comprising an additional active agent, such as an anti-cancer agent or an anti-angiogenic agent.

[0022] In certain aspects, the disclosure provides methods for using a VEGFR-2 binding protein to inhibit a VEGF biological activity in a cell or to inhibit a biological activity mediated by VEGFR-2. The cell may be situated in vivo or ex vivo, and may be, for example, a cell of a living organism, a cultured cell or a cell in a tissue sample. The method may comprise contacting said cell with any of the VEGFR-2-inhibiting polypeptides disclosed herein, in an amount and for a time sufficient to inhibit such biological activity.

[0023] In certain aspects, the disclosure provides methods for treating a subject having a condition which responds to the inhibition of VEGF or VEGFR-2. Such a method may comprise administering to said subject an effective amount of any of the VEGFR-2 inhibiting polypeptides described herein. A condition may be one that is characterized by inappropriate angiogenesis. A condition may be a hyperproliferative condition. Examples of conditions (or disorders) suitable for treatment include autoimmune disorders, inflammatory disorders, retinopathies (particularly proliferative retinopathies), and cancers. Any of the VEGFR-2 inhibiting polypeptides described herein may be used for the preparation of a medicament for the treatment of a disorder, particularly a disorder selected from the group consisting of: an autoimmune disorder, an inflammatory disorder, a retinopathy, and a cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

[0024] FIGS. 1A-1D are graphs and images depicting the characterization of KDR-binding single clones from Round 6 of KDR selection. FIG. 1A is a graph showing the specific binding of fibronectin-based binding proteins to 25 nM of KDR-Fc analyzed in radioactive equilibrium binding assay. FIG. 1B is a graph showing the inhibition of specific binding of KDR-Fc and selected fibronectin based binding proteins in the presence of 100-fold excess of VEGF.sub.165. As shown in this figure, certain binding proteins bound KDR-Fc competitively with VEGF.sub.165 while others, exemplified by clone 8, did not compete with VEGF.sub.165. FIG. 1C is a graph showing the inhibition of KDR-Fc interaction with immobilized VEGF.sub.165 in presence of selected fibronectin based binding proteins analyzed in BIAcore. FIG. 1D is an image showing binding of VR28 to KDR-expressing and control cells detected by immunofluorescence.

[0025] FIG. 2 is a graph showing the selection profile for the affinity maturation of VR28 KDR binder. Shown at left is binding of the VR28 clone to KDR-Fc and Flk1-Fc (very low, unlabeled bar). Shown at center is binding of a crude mutagenized pool and subsequent enrichment rounds to KDR-Fc. Shown at right is binding of further enrichment rounds to Flk-1-Fc. Binding was estimated in radioactive equilibrium binding assay as a percentage of input, using 1 nM KDR-Fc or Flk1-Fc.

[0026] FIGS. 3A and 3B are graphs depicting the characterization of KDR-binding single clones from Round 4 of anti-KDR affinity maturation of VR28 binder. FIG. 3A shows the saturation binding of VR28 (-.box-solid.-) and affinity matured K1 (-.tangle-solidup.-), K6 (--), K9 (-.diamond-solid.-), K10 (- -), K12 K13 (-.DELTA.-), K14 (-.gradient.-), K15 (-.diamond.-) to KDR-Fc in radioactive equilibrium binding assay FIG. 3B shows the binding of clones with and without N-terminal deletion to KDR-Fc. Deletion .DELTA.1-8 in the N-terminus of fibronectin-based binding proteins improved binding to KDR-Fc. The data represents an average KDR-Fc binding of 23 independent clones with and without N-terminal deletion.

[0027] FIG. 4 is a graph showing the binding of the selected clones to KDR and Flk-1. Specific binding of VR28 and selected clones after four rounds of affinity maturation to human KDR (K clones) and seven rounds of affinity maturation to human (KDR) and mouse (flk-1) (E clones). VEGFR-2-Fc chimeras were compared in radioactive equilibrium binding assay. The data represents an average of 3 independent experiments. As shown here, maturation against both mouse and human VEGFR-2 proteins produces binders that bind to both proteins.

[0028] FIGS. 5A and 5B are graphs showing the characterization of VEGFR-2-binding single clones from Round 7 of affinity maturation of VR28 binder. Saturation binding of VR28 (-.box-solid.-) and specificity matured E3 (-.tangle-solidup.-), E5 (--), E6 (-.diamond-solid.-), E9 (- -), E18 E19 (-.DELTA.-), E25 (-.gradient.-), E26 (-.diamond.-), E28 (-.largecircle.-), E29 (-X-) clones to KDR (FIG. 5A) and Flk1 (FIG. 5B)-Fc chimeras was tested in radioactive equilibrium binding assay.

[0029] FIGS. 6A and 6B are graphs showing the characterization of VEGFR-2 binding by single clones from Round 7 of affinity maturation of the VR28 binder. FIG. 6A shows the importance of arginine at positions 79 and 82 in binders with dual specificity to human and mouse VEGFR-2 for binding to mouse VEGFR-2 (Flk1). When either of these positions was replaced by amino acid other than R (X79=E, Q, W, P; X82=L, K), binding to Flk1 but not to KDR significantly decreased. FIG. 6B shows the importance of all three variable loops (BC, DE and FG) of KDR fibronectin-based binding proteins for binding to the target in these proteins. Substitution of each loop at a time by NNS sequence affected binding to KDR and Flk1. The binding data is an average from E6 and E26 clones.

[0030] FIGS. 7A and 7B are graphs showing the binding of selected fibronectin-based binding proteins to CHO cells expressing human KDR receptor (FIG. 7A) and EpoR-Flk1 chimera (FIG. 7B). E18 (-.box-solid.-), E19 (-.tangle-solidup.-), E26 (--), E29 (-.diamond-solid.-) and WT fibronectin-based scaffold proteins were tested. No binding to control CHO cells was observed (data not shown).

[0031] FIGS. 8A and 8B are graphs showing the inhibition of VEGF-induced proliferation of Ba/F3-KDR (FIG. 8A) and Ba/F3-Flk1 (FIG. 8B) cells, expressing KDR and Flk1 in the presence of different amounts of fibronectin-based binding proteins: E18 (-.box-solid.-), E19 (-.tangle-solidup.-), E26 (-.tangle-solidup.-), E29 (-.diamond-solid.-), M5 (- -), WT and anti-KDR or anti-flk-1 Ab (-.DELTA.-). The data represents an average of 2 independent experiments.

[0032] FIG. 9 is a graph showing the results of a HUVEC proliferation assay in the presence of different amounts of fibronectin-based scaffold proteins: E18 (-.box-solid.-), E19 (-.tangle-solidup.-), E26 (--), E29 (-.diamond-solid.-), M5 (- -), WT The data represents an average of 2 independent experiments. As shown, the KDR binding proteins caused a decrease in proliferation by approximately 40%.

[0033] FIG. 10 is a set of graphs showing the reversible refolding of M5FL in optimized buffer.

[0034] FIG. 11 is an image showing SDS-PAGE analysis of pegylated forms of M5FL. M, molecular weight markers [Sea Blue Plus, Invitrogen]; -, M5FL alone; 20, M5FL with 20 kD PEG; 40, M5FL with 40 kD PEG.

[0035] FIG. 12 is a graph showing the inhibition of VEGF-induced proliferation of Ba/F3-KDR cells with differing amounts of M5FL (-.diamond-solid.-), M5FL PEG20 (-.box-solid.-) and M5FL PEG40(-.tangle-solidup.-), respectively. Pegylation has little or no effect on M5FL activity in this assay.

[0036] FIG. 13 shows western analysis of VEGFR-2 signaling in endothelial cells. Phospho VEGFR-2--Visualization of phosphorylated VEGFR-2. VEGFR-2--Sample loading control. Phospho ERK1/2--Visualization of phosphorylated ERK1/2 (MAPK). ERK1-Sample loading control. The results demonstrated that 130 pM CT-01 blocks VEGFR-2 activation and signaling by VEGF-A.

[0037] FIG. 14 shows that various .sup.10Fn3-derived molecules (e.g. M5FL, F10, CT-01) can block VEGF-A and VEGF-D signaling.

[0038] FIG. 15 shows a comparison of .sup.125I native, pegylated CT-01 administered i.v. & i.p. CT-01 is a 12 kDa protein. It is rapidly cleared from the blood. Addition of a 40 kDa PEG reduces its clearance rate and increases the AUC by 10 fold. Half life of 16 hr in rats is equivalent to 2.times. dosing per week in humans. Administration route: i.p. CT-01-PEG40 has an AUC that is only 50% of an i.v. administration.

[0039] FIG. 16 shows the tissue distribution of .sup.125I-CT01PEG40 in normal rats. Tissue distribution of .sup.125I-CT01PEG40 indicates secretion primarily via the liver and secondarily via the kidney. This is expected for the high molecular weight PEG form. No long term accumulation of CT-01PEG40 is detected.

[0040] FIG. 17 is a schematic view of the Miles Assay that measures vascular permeability.

[0041] FIG. 18 shows that CT-01 blocks VEGF in vivo using the Miles Assay. The results indicate that 5 mg/kg of CT01-PEG40 blocks VEGF challenge.

[0042] FIG. 19 shows that CT-01 inhibits tumor growth using the B16-F10 Murine Melanoma Tumor Assay.

[0043] FIG. 20 shows that CT-01 inhibits tumor growth using U87 Human Glioblastoma.

[0044] FIGS. 21A and 21B show the sequences of VEGFR binding polypeptides that are based on an antibody light chain framework/scaffold (SEQ ID NOs:241-310).

[0045] FIG. 22 shows the structural organization for a single domain polypeptide having an immunoglobulin fold (a V.sub.H domain of an immunoglobulin, left side) and a single domain polypeptide having an immunoglobulin-like fold (a .sup.10Fn3 domain, right side).

DETAILED DESCRIPTION OF THE INVENTION

Definitions

[0046] A "functional Fc region" possesses at least one "effector function" of a native sequence Fc region. Exemplary "effector functions" include C1q binding; complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors (e.g. B cell receptor; BCR), etc. Such effector functions generally require the Fc region to be combined with a binding domain (e.g. an antibody variable domain) and can be assessed using various assays known in the art for evaluating such antibody effector functions.

[0047] A "native sequence Fc region" comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature.

[0048] A "variant Fc region" comprises an amino acid sequence which differs from that of a native sequence Fc region by virtue of at least one amino acid modification. Preferably, the variant Fc region has at least one amino acid substitution compared to a native sequence Fc region or to the Fc region of a parent polypeptide, e.g. from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions in a native sequence Fc region or in the Fc region of the parent polypeptide. The variant Fc region herein will preferably possess at least about 80% sequence identity with a native sequence Fc region and/or with an Fc region of a parent polypeptide, and most preferably at least about 90% sequence identity therewith, more preferably at least about 95% sequence identity therewith.

[0049] "Antibody-dependent cell-mediated cytotoxicity" and "ADCC" refer to a cell-mediated reaction in which nonspecific cytotoxic cells that express Fc receptors (FcRs) (e.g. Natural Killer (NK) cells, neutrophils, and macrophages) recognize bound antibody on a target cell and subsequently cause lysis of the target cell. The primary cells for mediating ADCC, NK cells, express Fc.gamma.RIII only, whereas monocytes express Fc.gamma.RI, Fc.gamma.RII and Fc.gamma.RIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991). To assess ADCC activity of a molecule of interest, an in vitro ADCC assay, such as that described in U.S. Pat. No. 5,500,362 or 5,821,337 may be performed. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells. Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in a animal model such as that disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998).

[0050] "Human effector cells" are leukocytes which express one or more FcRs and perform effector functions. Preferably, the cells express at least Fc.gamma.RIII and perform ADCC effector function. Examples of human leukocytes which mediate ADCC include peripheral blood mononuclear cells (PBMC), natural killer (NK) cells, monocytes, cytotoxic T cells and neutrophils; with PBMCs and NK cells being preferred. The effector cells may be isolated from a native source thereof, e.g. from blood or PBMCs as described herein.

[0051] The terms "Fc receptor" and "FcR" are used to describe a receptor that binds to the Fc region of an antibody. The preferred FcR is a native sequence human FcR. Moreover, a preferred FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the Fc.gamma.RI, Fc.gamma.RII, and Fc.gamma.RIII subclasses, including allelic variants and alternatively spliced forms of these receptors. Fc.gamma.RII receptors include Fc.gamma.RIIA (an "activating receptor") and Fc.gamma.RIIB (an "inhibiting receptor"), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof. Activating receptor Fc.gamma.RIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. Inhibiting receptor Fc.gamma.RIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (reviewed in Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein. The term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976); and Kim et al., J. Immunol. 24:249 (1994)).

[0052] "Percent (%) amino acid sequence identity" herein is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in a selected sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full-length of the sequences being compared. For purposes herein, however, % amino acid sequence identity values are obtained as described below by using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was authored by Genentech, Inc. has been filed with user documentation in the U.S. Copyright Office, Washington D.C., 20559, where it is registered under U.S. Copyright Registration No. TXU510087, and is publicly available through Genentech, Inc., South San Francisco, Calif. The ALIGN-2 program should be compiled for use on a UNIX operating system, preferably digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary.

[0053] For purposes herein, the % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (which can alternatively be phrased as a given amino acid sequence A that has or comprises a certain % amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows: 100 times the fraction X/Y where X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 in that program's alignment of A and B, and where Y is the total number of amino acid residues in B. It will be appreciated that where the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not equal the % amino acid sequence identity of B to A.

[0054] A "polypeptide chain" is a polypeptide wherein each of the domains thereof is joined to other domain(s) by peptide bond(s), as opposed to non-covalent interactions or disulfide bonds.

[0055] An "isolated" polypeptide is one that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In preferred embodiments, the polypeptide will be purified (1) to greater than 95% by weight of polypeptide as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain. Isolated polypeptide includes the polypeptide in situ within recombinant cells since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step.

[0056] Targets may also be fragments of said targets. Thus a target is also a fragment of said target, capable of eliciting an immune response. A target is also a fragment of said target, capable of binding to a single domain antibody raised against the full length target.

[0057] A fragment as used herein refers to less than 100% of the sequence (e.g., 99%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% etc.), but comprising 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more amino acids. A fragment is of sufficient length such that the interaction of interest is maintained with affinity of 1.times.10.sup.-6M or better.

[0058] A fragment as used herein also refers to optional insertions, deletions and substitutions of one or more amino acids which do not substantially alter the ability of the target to bind to a single domain antibody raised against the wild-type target. The number of amino acid insertions deletions or substitutions is preferably up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69 or 70 amino acids.

[0059] A protein of the invention that "induces cell death" is one which causes a viable cell to become nonviable. The cell is generally one which expresses the antigen to that the protein binds, especially where the cell overexpresses the antigen. Preferably, the cell is a cancer cell, e.g. a breast, ovarian, stomach, endometrial, salivary gland, lung, kidney, colon, thyroid, pancreatic or bladder cell. In vitro, the cell may be a SKBR3, BT474, Calu 3, MDA-MB453, MDA-MB-361 or SKOV3 cell. Cell death in vitro may be determined in the absence of complement and immune effector cells to distinguish cell death induced by antibody dependent cell-mediated cytotoxicity (ADCC) or complement dependent cytotoxicity (CDC). Thus, the assay for cell death may be performed using heat inactivated serum (i.e. in the absence of complement) and in the absence of immune effector cells. To determine whether the protein of the invention is able to induce cell death, loss of membrane integrity as evaluated by uptake of propidium iodide (PI), trypan blue (see Moore et al. Cytotechnology 17:1-11 (1995)) or 7AAD can be assessed relative to untreated cells.

[0060] A protein of the invention that "induces apoptosis" is one that induces programmed cell death as determined by binding of apoptosis related molecules or events, such as annexin V, fragmentation of DNA, cell shrinkage, dilation of endoplasmic reticulum, cell fragmentation, and/or formation of membrane vesicles (called apoptotic bodies). The cell is one which expresses the antigen to which the protein binds and may be one which overexpresses the antigen. The cell may be a tumor cell, e.g. a breast, ovarian, stomach, endometrial, salivary gland, lung, kidney, colon, thyroid, pancreatic or bladder cell. In vitro, the cell may be a SKBR3, BT474, Calu 3 cell, MDA-MB453, MDA-MB-361 or SKOV3 cell. Various methods are available for evaluating the cellular events associated with apoptosis. For example, phosphatidyl serine (PS) translocation can be measured by annexin binding; DNA fragmentation can be evaluated through DNA laddering as disclosed in the example herein; and nuclear/chromatin condensation along with DNA fragmentation can be evaluated by any increase in hypodiploid cells. Preferably, the protein that induces apoptosis is one which results in about 2 to 50 fold, preferably about 5 to 50 fold, and most preferably about 10 to 50 fold, induction of annexin binding relative to untreated cell in an annexin binding assay using cells expressing the antigen to which the protein of the invention binds.

[0061] The term "therapeutically effective amount" refers to an amount of a drug effective to treat a disease or disorder in a mammal. In the case of cancer, the therapeutically effective amount of the drug may reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; and/or relieve to some extent one or more of the symptoms associated with the disorder. To the extent the drug may prevent growth and/or kill existing cancer cells, it may be cytostatic and/or cytotoxic. For cancer therapy, efficacy in vivo can, for example, be measured by assessing the time to disease progression (TTP) and/or determining the response rates (RR).

[0062] The term "PK" is an acronym for "pharmokinetic" and encompasses properties of a compound including, by way of example, absorbtion, distribution, metabolism, and elimination by a subject. A "PK modulation protein" refers to any protein or peptide that affects the pharmokinetic properties of a biologically active molecule when fused to or administered together with the biologically active molecule. Examples of a PK modulation protein include PEG, as well as human serum albumin (HSA) binders as disclosed in US Pat. App. Nos. 20050287153 and 20070003549.

[0063] As used herein, "ocular" refers to the eye, surrounding tissues, and to bodily fluids in the region of the eye. Specifically, the term includes the cornea, the sclera, the uvea, the conjunctiva (e.g., bulbar conjunctiva, palpebral conjunctiva, and tarsal conjunctiva), anterior chamber, lacrimal sac, lacrimal canals, lacrimal ducts, medial canthus, nasolacrimal duct, and the eyelids (e.g., upper eyelid and lower eyelid). Additionally, the term includes the inner surface of the eye (conjunctiva overlying the sclera), and the inner surface of the eyelids (palpepral conjunctiva).

Overview

[0064] The present application provides novel sustained-release intraocular drug delivery systems that are particularly useful in treating disorders of the eye. In exemplary embodiments, the drug delivery systems deliver VEGFR-2 specific inhibitors intraocularly.

[0065] In one aspect, sustained-release intraocular drug delivery system is provided comprising a therapeutic component and a polymeric component. The therapeutic component comprises an antiangiogenic polypeptide component such as an antibody, an antibody fragment, or an artificial antibody, as well as humanized versions.

Polymeric Component

[0066] In some embodiments, the polymeric component of the sustained-release drug delivery system comprises monomers such as organic esters or ethers, which when degraded result in physiologically acceptable degradation products. Anhydrides, amides, orthoesters, or the like, by themselves or in combination with other monomers, may also be used. The polymers are generally condensation polymers. The polymers can be crosslinked or non-crosslinked. If crosslinked, they are usually not more than lightly crosslinked, and are less than 5% crosslinked, usually less than 1% crosslinked.

[0067] In addition to carbon and hydrogen, the polymers will include oxygen and nitrogen, particularly oxygen. The oxygen may be present as oxy, e.g., hydroxy or ether, carbonyl, e.g., non-oxo-carbonyl, such as carboxylic acid ester, and the like. The nitrogen can be present as amide, cyano, and amino. An exemplary list of biodegradable polymers that can be used are described in Heller, Biodegradable Polymers in Controlled Drug Delivery, In: "CRC Critical Reviews in Therapeutic Drug Carrier Systems," Vol. 1. CRC Press, Boca Raton, Fla. (1987).

[0068] Of particular interest are polymers of hydroxyaliphatic carboxylic acids, either homo- or copolymers, and polysaccharides. Included among the polyesters of interest are homo- or copolymers of D-lactic acid, L-lactic acid, racemic lactic acid, glycolic acid, caprolactone, and combinations thereof. Copolymers of glycolic and lactic acid are of particular interest, where the rate of biodegradation is controlled by the ratio of glycolic to lactic acid. The percent of each monomer in poly(lactic-co-glycolic)acid (PLGA) copolymer may be 0-100%, about 15-85%, about 25-75%, or about 35-65%. In certain variations, 25/75 PLGA and/or 50/50 PLGA copolymers are used. In other variations, PLGA copolymers are used in conjunction with polylactide polymers.

[0069] Biodegradable polymer matrices that include mixtures of hydrophilic and hydrophobic ended PLGA may also be employed, and are useful in modulating polymer matrix degradation rates. Hydrophobic ended (also referred to as capped or end-capped) PLGA has an ester linkage hydrophobic in nature at the polymer terminus. Typical hydrophobic end groups include, but are not limited to alkyl esters and aromatic esters. Hydrophilic ended (also referred to as uncapped) PLGA has an end group hydrophilic in nature at the polymer terminus. PLGA with a hydrophilic end groups at the polymer terminus degrades faster than hydrophobic ended PLGA because it takes up water and undergoes hydrolysis at a faster rate (Tracy et al., Biomaterials 20: 1057-1062 (1999)). Examples of suitable hydrophilic end groups that may be incorporated to enhance hydrolysis include, but are not limited to, carboxyl, hydroxyl, and polyethylene glycol. The specific end group will typically result from the initiator employed in the polymerization process. For example, if the initiator is water or carboxylic acid, the resulting end groups will be carboxyl and hydroxyl. Similarly, if the initiator is a monofunctional alcohol, the resulting end groups will be ester or hydroxyl.

[0070] Further polymers and polymer blends useful in the invention can be found in U.S. Patent Applications 20060210604, 20070088014, and 20070059336 hereby incorporated by reference.

Antiangiogenic Polypeptides

[0071] In some embodiments, the antiangiogenic polypeptide component comprises one or more single domain polypeptides that may be derived from two related groups of protein structures: those proteins having an immunoglobulin fold, such as an antibody, and those proteins having an immunoglobulin-like fold, such as an artificial antibody. "Artificial antibody" is meant to include fibronectin based scaffold proteins such as the Adnectins.TM. or the "addressable" therapeutic binding molecules. By a "polypeptide" is meant any sequence of two or more amino acids, regardless of length, post-translation modification, or function. "Polypeptide," "peptide," and "protein" are used interchangeably herein. Polypeptides can include natural amino acids and non-natural amino acids such as those described in U.S. Pat. No. 6,559,126, incorporated herein by reference. Polypeptides can also be modified in any of a variety of standard chemical ways (e.g., an amino acid can be modified with a protecting group; the carboxy-terminal amino acid can be made into a terminal amide group; the amino-terminal residue can be modified with groups to, e.g., enhance lipophilicity; or the polypeptide can be chemically glycosylated or otherwise modified to increase stability or in vivo half-life). Polypeptide modifications can include the attachment of another structure such as a cyclic compound or other molecule to the polypeptide and can also include polypeptides that contain one or more amino acids in an altered configuration (i.e., R or S; or, L or D). The term "single domain polypeptide" is used to indicate that the target binding activity (e.g., VEGFR-2 binding activity) of the subject polypeptide is situated within a single structural domain, as differentiated from, for example, antibodies and single chain antibodies, where antigen binding activity is generally contributed by both a heavy chain variable domain and a light chain variable domain. It is contemplated that a plurality of single domain polypeptides of the sort disclosed herein could be connected to create a composite molecule with increased avidity. Likewise, a single domain polypeptide may be attached (e.g., as a fusion protein) to any number of other polypeptides, such as fluorescent polypeptides, targeting polypeptides and polypeptides having a distinct therapeutic effect.

[0072] Single domain polypeptides of either the immunoglobulin or immunoglobulin-like scaffold will tend to share certain structural features. For example, the polypeptide may comprise between about 80 and about 150 amino acids, which amino acids are structurally organized into a set of beta or beta-like strands, forming beta sheets, where the beta or beta-like strands are connected by intervening loop portions. The beta sheets form the stable core of the single domain polypeptides, while creating two "faces" composed of the loops that connect the beta or beta-like strands. As described herein, these loops can be varied to create customized ligand binding sites, and, with proper control, such variations can be generated without disrupting the overall stability of the protein. In antibodies, three of these loops are the well-known Complementarity Determining Regions (or "CDRs").

[0073] Scaffolds for formation of a single domain polypeptides should be highly soluble and stable in physiological conditions. Examples of immunoglobulin scaffolds are the single domain V.sub.H or V.sub.L scaffold, as well as a single domain camelid V.sub.HH domain (a form of variable heavy domain found in camelids) or other immunoglobulin variable domains found in nature or engineered in the laboratory. In the single domain format disclosed herein, an immunoglobulin polypeptide need not form a dimer with a second polypeptide in order to achieve binding activity. Accordingly, any such polypeptides that naturally contain a cysteine which mediates disulfide cross-linking to a second protein can be altered to eliminate the cysteine. Alternatively, the cysteine may be retained for use in conjugating additional moieties, such as PEG, to the single domain polypeptide.

[0074] Other scaffolds may be non-antibody scaffold proteins. By "non-antibody scaffold protein or domain" is meant a non-antibody polypeptide having an immunoglobulin-like fold. By "immunoglobulin-like fold" is meant a protein domain of between about 80-150 amino acid residues that includes two layers of antiparallel beta-sheets, and in which the flat, hydrophobic faces of the two beta-sheets are packed against each other. An example of such a scaffold is the "fibronectin-based scaffold protein", by which is meant a polypeptide based on a fibronectin type III domain (Fn3). Fibronectin is a large protein which plays essential roles in the formation of extracellular matrix and cell-cell interactions; it consists of many repeats of three types (types I, II, and III) of small domains (Baron et al., 1991). Fn3 itself is the paradigm of a large subfamily which includes portions of cell adhesion molecules, cell surface hormone and cytokine receptors, chaperoning, and carbohydrate-binding domains for reviews, see Bork & Doolittle, Proc Natl Acad Sci USA. 1992 Oct. 1; 89(19):8990-4; Bork et al., J Mol Biol. 1994 Sep. 30; 242(4):309-20; Campbell & Spitzfaden, Structure. 1994 May 15; 2(5):333-7; Harpez & Chothia, J Mol Biol. 1994 May 13; 238(4):528-39).

[0075] Preferably, the fibronectin-based scaffold protein is a ".sup.10FN3" scaffold, by which is meant a polypeptide variant based on the tenth module of the human fibronectin type III protein in which one or more of the solvent accessible loops has been randomized or mutated, particularly one or more of the three loops identified as the BC loop (amino acids 23-30), DE loop (amino acids 52-56) and FG loop (amino acids 77-87) (the numbering scheme is based on the sequence on the tenth Type III domain of human fibronectin, with the amino acids Val-Ser-Asp-Val-Pro representing amino acids numbers 1-5). The amino acid sequence of the wild-type tenth module of the human fibronectin type III domain is:

VSDVPRDLEVVAATPTSLLISWDAPAVTVRYYRITYGETGGNSPVQEFTVPGSKS TATISGLKPGVDYTITGYAVTGRGDSPASSKPISINYRT (SEQ ID NO:5). Thus, the wild-type BC loop comprises the sequence of DAPAVTVR; the wild-type DE loop comprises the sequence of GSKST; the wild-type FG loop comprises the sequence of GRGDSPASSKP.

[0076] A variety of improved mutant .sup.10Fn3 scaffolds have been identified. A modified Asp7, which is replaced by a non-negatively charged amino acid residue (e.g., Asn, Lys, etc.). Both of these mutations have the effect of promoting greater stability of the mutant .sup.10Fn3 at neutral pH as compared to the wild-type form. A variety of additional alterations in the .sup.10Fn3 scaffold that are either beneficial or neutral have been disclosed. See, for example, Batori et al. Protein Eng. 2002 December; 15(12):1015-20; Koide et al., Biochemistry 2001 Aug. 28; 40(34):10326-33.

[0077] Both the variant and wild-type .sup.10Fn3 proteins are characterized by the same structure, namely seven beta-strand domain sequences (designated A through and six loop regions (AB loop, BC loop, CD loop, DE loop, EF loop, and FG loop) which connect the seven beta-strand domain sequences. The beta strands positioned closest to the N- and C-termini may adopt a beta-like conformation in solution. In SEQ ID NO:5, the AB loop corresponds to residues 15-16, the BC loop corresponds to residues 22-30, the CD loop corresponds to residues 39-45, the DE loop corresponds to residues 51-55, the EF loop corresponds to residues 60-66, and the FG loop corresponds to residues 76-87. The BC loop, DE loop, and FG loop are all located at the same end of the polypeptide. Similarly, immunoglobulin scaffolds tend to have at least seven beta or beta-like strands, and often nine beta or beta-like strands. Adnectins.TM. can include other Fn3 type fibronectin domains as long as they exhibit useful activities and properties of .sup.10Fn3 type domains.

VEGFR-2 Binding Proteins

[0078] In preferred embodiments of the invention, the antiangiogenic polypeptide component comprises a single domain polypeptide that is a VEGFR-2 specific binder. A single domain polypeptide disclosed herein may have at least five to seven beta or beta-like strands distributed between at least two beta sheets, and at least one loop portion connecting two beta or beta-like strands, which loop portion participates in binding to VEGFR-2, particularly KDR, with the binding characterized by a dissociation constant that is less than 1.times.10.sup.-6M, and preferably less than 1.times.10.sup.-8M. As described herein, polypeptides having a dissociation constant of less than 5.times.10.sup.-9M are particularly desirable for therapeutic use in vivo to inhibit VEGF signaling. Polypeptides having a dissociation constant of between 1.times.10.sup.-6 M and 5.times.10.sup.-9M may be desirable for use in detecting or labeling, ex vivo or in vivo, VEGFR-2 proteins.

[0079] Optionally, the VEGFR-2 binding protein will bind specifically to VEGFR-2 relative to other related proteins from the same species. By "specifically binds" is meant a polypeptide that recognizes and interacts with a target protein (e.g., VEGFR-2) but that does not substantially recognize and interact with other molecules in a sample, for example, a biological sample. In preferred embodiments a polypeptide of the invention will specifically bind a VEGFR with a K.sub.D at least as tight as 500 nM. Preferably, the polypeptide will specifically bind a VEGFR with a K.sub.D of 1 pM to 500 nM, more preferably 1 pM to 100 nM, more preferably 1 pM to 10 nM, and most preferably 1 pM to 1 nM or lower.

[0080] In general, a library of scaffold single domain polypeptides is screened to identify specific polypeptide variants that can bind to a chosen target. These libraries may be, for example, phage display libraries or PROfusion.TM. libraries.

[0081] In an exemplary embodiment, we have exploited a novel in vitro RNA-protein fusion display technology to isolate polypeptides that bind to both human (KDR) and mouse (Flk-1) VEGFR-2 and inhibit VEGF-dependent biological activities. These polypeptides were identified from libraries of fibronectin-based scaffold proteins (Koide et al, JMB 284:1141 (1998)) and libraries of V.sub.L domains in which the diversity of CDR3 has been increased by swapping with CDR3 domains from a population of V.sub.H molecules. .sup.10Fn3 comprises approximately 94 amino acid residues, as shown in SEQ ID NO:5.

[0082] In addition, as described above, amino acid sequences at the N-terminus of .sup.10Fn3 can also be mutated or deleted. For example, randomization of the BC, DE, and FG loops can occur in the context of a full-length .sup.10Fn3 or in the context of a .sup.10Fn3 having a deletion or mutation of 1-8 amino acids of the N-terminus. For example, the L at position 8 can be mutated to a Q. After randomization to create a diverse library, fibronectin-based scaffold proteins can be used in a screening assay to select for polypeptides with a high affinity for a protein, in this case the VEGFR. (For a detailed description of the RNA-protein fusion technology and fibronectin-based scaffold protein library screening methods see Szostak et al., U.S. Pat. Nos. 6,258,558; 6,261,804; 6,214,553; 6,281,344; 6,207,446; 6,518,018; PCT Publication Numbers WO 00/34784; WO 01/64942; WO 02/032925; and Roberts and Szostak, Proc Natl. Acad. Sci. 94:12297-12302, 1997, herein incorporated by reference.)

[0083] For the initial selection described herein, three regions of the .sup.10Fn3 at positions 23-29, 52-55 and 77-86 were randomized and used for in vitro selection against the extracellular domain of human VEGFR-2 (amino acids 1-764 of KDR fused to human IgG1Fc). Using mRNA display (RNA-protein fusion) and in vitro selection, we sampled a .sup.10Fn3-based library with approximately ten trillion variants. The initial selection identified polypeptides with moderate affinity (K.sub.D=0-200 nM) that competed with VEGF for binding to KDR (human VEGFR-2). Subsequently, a single clone (K.sub.D=11-13 nM) from the initial selection was subjected to mutagenesis and further selection. This affinity maturation process yielded new VEGFR binding polypeptides with dissociation constants between 60 pM to 2 nM. KDR binders are shown in Table 3. In addition, we also isolated polypeptides that could bind to Flk-1, the mouse KDR homolog, from mutagenized populations of KDR binders that initially had no detectable binding affinity to Flk-1, resulting in the isolation of polypeptides that exhibit dual specificities to both human and mouse VEGFR-2. These polypeptides are shown to bind cells that display KDR or Flk-1 extracellular domains. They also inhibited cell growth in a VEGF-dependent proliferation assay. Polypeptides that bind to KDR and Flk-1 are shown in Table 2, while a selection of preferred KDR binders and KDR/Flk-1 binders are shown in Table 1.

[0084] Using the VEGFR-2 binding polypeptides identified in these selections we determined FG loop amino acid consensus sequences required for the binding of the polypeptides to the VEGFR-2. The sequences are listed as SEQ ID NOs:1-4 below.

[0085] VEGFR-2 binding polypeptides, such as those of SEQ ID NOs:1-4, may be formulated alone (as isolated peptides), as part of a .sup.10Fn3 single domain polypeptide, as part of a full-length fibronectin, (with a full-length amino terminus or a deleted amino terminus) or a fragment thereof, in the context of an immunoglobulin (particularly a single domain immunoglobulin), in the context of another protein having an immunoglobulin-like fold, or in the context of another, unrelated protein. The polypeptides can also be formulated as part of a fusion protein with a heterologous protein that does not itself bind to or contribute in binding to a VEGFR. In addition, the polypeptides of the invention can also be fused to nucleic acids. The polypeptides can also be engineered as monomers, dimers, or multimers.

[0086] Sequences of the Preferred Consensus VEGFR-2 Binding Peptides:

TABLE-US-00001 SEQ ID NO:1- (L/M)GXN(G/D)(H/R)EL(L/M)TP

[X can be any amino acid; (/) represents alternative amino acid for the same position]

TABLE-US-00002 SEQ ID NO:2- XERNGRXL(L/M/N)TP

[X can be any amino acid; (/) represents alternative amino acid for the same position]

TABLE-US-00003 SEQ ID NO:3- (D/E)GXNXRXXIP

[X can be any amino acid; (/) represents alternative amino acid for the same position]

TABLE-US-00004 SEQ ID NO:4- (D/E)G(R/P)N(G/E)R(S/L)(S/F)IP

[X can be any amino acid; (/) represents alternative amino acid for the same position]

[0087] Sequences of the Preferred VEGFR-2 Binding .sup.10Fn3 Polypeptides:

TABLE-US-00005 SEQ ID NO:6 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTMGLYGHELLTPISTNYRT SEQ ID NO:7 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTDGENGQFLLVPISINYRT SEQ ID NO:8 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTMGPNDNELLTPISINYRT SEQ ID NO:9 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTAGWDDHELFIPISINYRT SEQ ID NO:10 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTSGHNDHMLMIPISINYRT SEQ ID NO:11 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTAGYNDQILMTPISINYRT SEQ ID NO:12 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTFGLYGKELLIPISINYRT SEQ ID NO:13 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTTGPNDRLLFVPISINYRT SEQ ID NO:14 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTDVYNDHEIKTPISINYRT SEQ ID NO:15 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTDGKDGRVLLTPISINYRT SEQ ID NO:16 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTEVHHDREIKTPISINYRT SEQ ID NO:17 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTQAPNDRVLYTPISINYRT SEQ ID NO:18 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTREENDHELLIPISINYRT SEQ ID NO:19 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTVTHNGHPLMTPISINYRT SEQ ID NO:20 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTLALKGHELLTPISINYRT SEQ ID NO:21 VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPPTATISGLKPGVDYTITGYAVTVAQNDHELITPISINYRT SEQ ID NO:22 VSDVPRDL/QEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEF TVPLQPPAATISGLKPGVDYTITGYAVTMAQSGHELFTPISINYRT SEQ ID NO:24 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTVERNGRVLMTPISINYRT SEQ ID NO:25 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTVERNGRHLMTPISINYRT SEQ ID NO:33 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTLERNGRELMTPISINYRT SEQ ID NO:45 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTEERNGRTLRTPISINYRT SEQ ID NO:53 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTVERNDRVLFTPISINYRT SEQ ID NO:57 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTVERNGRELMTPISINYRT SEQ ID NO:62 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTLERNGRELMVPISINYRT SEQ ID NO:63 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTDGRNDRKLMVPISINYRT SEQ ID NO:68 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTDGQNGRLLNVPISINYRT SEQ ID NO:91 EVVAATPTSLLISWRHHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTA TISGLKPGVDYTITGYAVTVHWNGRELMTPISINYRT SEQ ID NO:92 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTEEWNGRVLMTPISINYRT SEQ ID NO:93 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTVERNGHTLMTPISINYRT SEQ ID NO:94 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTVEENGRQLMTPISINYRT SEQ ID NO:95 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTLERNGQVLFTPISINYRT SEQ ID NO:96 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTVERNGQVLYTPISINYRT SEQ ID NO:97 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTWGYKDHELLIPISINYRT SEQ ID NO:98 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTLGRNDRELLTPISINYRT SEQ ID NO:99 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTDGPNDRLLNIPISINYRT SEQ ID NO:100 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTFARDGHEILTPISINYRT SEQ ID NO:101 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTLEQNGRELMTPISINYRT SEQ ID NO:102 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTVEENGRVLNTPISINYRT SEQ ID NO:103 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTLEPNGRYLMVPISINYRT SEQ ID NO:104 EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITGYAVTEGRNGRELFIPISINYRT SEQ ID NO:154 VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPPAATISGLKPGVDYTITGYAVTWERNGRELFTPISINYRT SEQ ID NO:156 VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPPAATISGLKPGVDYTITGYAVTKERNGRELFTPISINYRT SEQ ID NO:172 VSDVPRDLEVVAATPTSLLISWRHPHFPTHYYRITYGETGGNSPVQEFTV PLQPPAATISGLKPGVDYTITGYAVTTERTGRELFTPISINYRT SEQ ID NO:173 VSDVPRDLEVVAATPTSLLISWRHPHFPTHYYRITYGETGGNSPVQEFTV PLQPPAATISGLKPGVDYTITGYAVTKERSGRELFTPISINYRT SEQ ID NO:175 VSDVPRDLEVVAATPTSLLISWRHPHFPTHYYRITYGETGGNSPVQEFTV PLQPPAATISGLKPGVDYTITGYAVTLERDGRELFTPISINYRT SEQ ID NO:177 VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPPLATISGLKPGVDYTITG/VYAVTKERNGRELFTPISINYRT SEQ ID NO:180 VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPTTATISGLKPGVDYTITGYAVTWERNGRELFTPISINYRT SEQ ID NO:181 VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPTVATISGLKPGVDYTITGYAVTLERNDRELFTPISINYRT SEQ ID NO:186 MGEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPT ATISGLKPGVDYTITVYAVTDGRNGRLLSIPISINYRTEIDKPSQ SEQ ID NO:187 MGEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPT ATISGLKPGVDYTITVYAVTDGRNGRLLSIPISINYRTEIDKPCQ

SEQ ID NO:188 MVSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFT VPLQPPTATISGLKPGVDYTITVYAVTDGRNGRLLSIPISINYRTEIDKPSQ SEQ ID NO:189 MGEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPT ATISGLKPGVDYTITVYAVTDGWNGRLLSIPISINYRT SEQ ID NO:190 MGEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPT ATISGLKPGVDYTITVYAVTEGPNERSLFIPISINYRT SEQ ID NO:191 MVSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFT VPLQPPTATISGLKPGVDYTITVYAVTEGPNERSLFIPISINYRT SEQ ID NO:192 (A core form of the polypeptide referred to herein as CT-01): EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITVYAVTDGRNGRLLSIPISINYRT

[0088] The CT-01 molecule above has a deletion of the first 8 amino acids and may include additional amino acids at the N- or C-termini. For example, an additional MG sequence may be placed at the N-terminus. The M will usually be cleaved off, leaving a GEV . . . sequence at the N-terminus. The re-addition of the normal 8 amino acids at the N-terminus also produces a KDR binding protein with desirable properties. The N-terminal methionine is generally cleaved off to yield a sequence:

TABLE-US-00006 (SEQ ID NO:193) VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPPTATISGLKPGVDYTITVYAVTDGRNGRLLSIPISINYRT.

[0089] A polypeptide disclosed herein may be modified by one or more conservative substitutions, particularly in portions of the protein that are not expected to interact with a target protein. It is expected that as many as 5%, 10%, 20% or even 30% or more of the amino acids in an immunoglobulin or immunoglobulin-like domain may be altered by a conservative substitution without substantially altering the affinity of the protein for target. It may be that such changes will alter the immunogenicity of the polypeptide in vivo, and where the immunogenicity is decreased, such changes will be desirable. As used herein, "conservative substitutions" are residues that are physically or functionally similar to the corresponding reference residues. That is, a conservative substitution and its reference residue have similar size, shape, electric charge, chemical properties including the ability to form covalent or hydrogen bonds, or the like. Preferred conservative substitutions are those fulfilling the criteria defined for an accepted point mutation in Dayhoff et al., Atlas of Protein Sequence and Structure 5:345-352 (1978 & Supp.). Examples of conservative substitutions are substitutions within the following groups: (a) valine, glycine; (b) glycine, alanine; (c) valine, isoleucine, leucine; (d) aspartic acid, glutamic acid; (e) asparagine, glutamine; (f) serine, threonine; (g) lysine, arginine, methionine; and (h) phenylalanine, tyrosine.

[0090] Polypeptides disclosed herein may also be modified in order to improve potency, bioavailability, chemical stability, and/or efficacy. For example, within one embodiment of the invention D-amino acid peptides, or retroenantio peptide sequences may be generated in order to improve the bioactivity and chemical stability of a polypeptide structure (see, e.g., Juvvadi et al., J. Am. Chem. Soc. 118: 8989-8997, 1996; Freidinger et al., Science, 210: 656-658, 1980). Lactam constraints (see Freidinger, supra), and/or azabicycloalkane amino acids as dipeptide surrogates can also be utilized to improve the biological and pharmacological properties of the native peptides (see, e.g., Hanessian et al., Tetrahedron 53:12789-12854, 1997).

[0091] Amide bond surrogates, such as thioamides, secondary and tertiary amines, heterocycles among others (see review in Spatola, A. F. in "Chemistry and Biochemistry of Amino Acids, Peptides and Proteins" Wenstein, B. Ed. Marcel Dekker, New York, 1983 Vol. 7, pp 267-357) can also be utilized to prevent enzymatic degradation of the polypeptide backbone thereby resulting in improved activity. Conversion of linear polypeptides to cyclic polypeptide analogs can also be utilized to improve metabolic stability, since cyclic polypeptides are much less sensitive to enzymatic degradation (see generally, Veber, et al. Nature 292:55-58, 1981).

[0092] Polypeptides can also be modified utilizing end group capping as esters and amides in order to slow or prevent metabolism and enhance lipophilicity. Dimers of the peptide attached by various linkers may also enhance activity and specificity (see for example: Y. Shimohigashi et al, in Peptide Chemistry 1988, Proceedings of the 26th Symposium on Peptide Chemistry, Tokyo, October 24-26, pgs. 47-50, 1989). For additional examples of polypeptide modifications, such as non-natural amino acids, see U.S. Pat. No. 6,559,126.

[0093] For use in vivo, a form suitable for pegylation may be generated. For example, a C-terminal tail comprising a cysteine was added and expressed, as shown below for a CT-01 form lacking the eight N-terminal amino acids (EIDKPCQ is added at the C-terminus).

GEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTATISGLK PGVDYTITVYAVTDGRNGRLLSIPISINYRTEIDKPCQ (SEQ ID NO:194). The pegylated form of this molecule is used in the in vivo experiments described below. A control form with a serine instead of a cysteine was also used:

TABLE-US-00007 (SEQ ID NO:195) GEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTA TISGLKPGVDYTITVYAVTDGRNGRLLSIPISINYRTEIDKPSQ.

[0094] The same C-terminal tails may also be added to CT-01 forms having the N-terminal eight amino acids, such as is shown in SEQ ID NO:193.

[0095] Additional variants with desirable KDR binding properties were isolated. The following core sequence has a somewhat different FG loop, and may be expressed with, for example, an N-terminal MG sequence, an N-terminal sequence that restores the 8 deleted amino acids, and/or a C-terminal tail to provide a cysteine for pegylation.

EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTATISGLKP GVDYTITVYAVTEGPNERSLFIPISINYRT (SEQ ID NO:196). Another such variant has the core sequence:

TABLE-US-00008 (SEQ ID NO:197) VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPPTATISGLKPGVDYTITVYAVTEGPNERSLFIPISTNYRT.

[0096] Additionally, preferred single domain immunoglobulin polypeptides in a V.sub.L framework were isolated by similar methodology and are disclosed in FIG. 21.

ADDITIONAL PROTEIN EMBODIMENTS

[0097] Proteins of the invention include a single domain polypeptide as described herein, generally a polypeptide that binds to a target, such as VEGFR-2, and where target binding activity situated within a single structural domain, as differentiated from, for example, antibodies and single chain antibodies, where antigen binding activity is generally contributed by both a heavy chain variable domain and a light chain variable domain. The disclosure also provides larger proteins that may comprise single domain polypeptides that bind to target. For example, a plurality of single domain polypeptides may be connected to create a composite molecule with increased avidity or multivalency. Likewise, a single domain polypeptide may be attached (e.g., as a fusion protein) to any number of other polypeptides. In certain aspects a single domain polypeptide may comprise at least five to seven beta or beta-like strands distributed among at least two beta sheets, as exemplified by immunoglobulin and immunoglobulin-like domains. A beta-like strand is a string of amino acids that participates in the stabilization of a single domain polypeptide but does not necessarily adopt a beta strand conformation. Whether a beta-like strand participates in the stabilization of the protein may be assessed by deleting the string or altering the sequence of the string and analyzing whether protein stability is diminished. Stability may be assessed by, for example, thermal denaturation and renaturation studies. Preferably, a single domain polypeptide will include no more than two beta-like strands. A beta-like strand will not usually adopt an alpha-helical conformation but may adopt a random coil structure. In the context of an immunoglobulin domain or an immunoglobulin-like domain, a beta-like strand will most often occur at the position in the structure that would otherwise be occupied by the most N-terminal beta strand or the most C-terminal beta strand. An amino acid string which, if situated in the interior of a protein sequence would normally form a beta strand, may, when situated at a position closer to an N- or C-terminus, adopt a conformation that is not clearly a beta strand and is referred to herein as a beta-like strand.

[0098] In certain embodiments, the disclosure provides single domain polypeptides that bind to VEGFR-2. Preferably the single domain polypeptides bind to human VEGFR-2 or a model species VEGFR-2. A single domain polypeptide may comprise between about 80 and about 150 amino acids that have a structural organization comprising: at least seven beta strands or beta-like strands distributed between at least two beta sheets, and at least one loop portion connecting two beta strands or beta-like strands, which loop portion participates in binding to VEGFR-2. In other words a loop portion may link two beta strands, two beta-like strands or one beta strand and one beta-like strand. Typically, one or more of the loop portions will participate in VEGFR-2 binding, although it is possible that one or more of the beta or beta-like strand portions will also participate in VEGFR-2 binding, particularly those beta or beta-like strand portions that are situated closest to the loop portions. A single domain polypeptide may comprise a structural unit that is an immunoglobulin domain or an immunoglobulin-like domain. A single domain polypeptide may bind to any part of VEGFR-2, although polypeptides that bind to an extracellular domain of a VEGFR-2 are preferred. Binding may be assessed in terms of equilibrium constants (e.g., dissociation, K.sub.D) and in terms of kinetic constants (e.g., on rate constant, k.sub.on and off rate constant, k.sub.off). A single domain polypeptide will typically be selected to bind to VEGFR-2 with a K.sub.D of less than about 10.sup.-6M, or less than about 10.sup.-7M, about 5.times.10.sup.-8M, about 10.sup.-8M or less than about 10.sup.-9M. VEGFR-2 binding polypeptides may compete for binding with one, or two or more members of the VEGF family, particularly VEGF-A, VEGF-C, and/or VEGF-D, and may inhibit one or more VEGFR-2-mediated biological events, such as proliferation of cancer cells and cancer metastasis. VEGFR-2 binding polypeptides may be used for therapeutic purposes as well as for any purpose involving the detection or binding of VEGFR-2. Polypeptides for therapeutic use will generally have a K.sub.D of less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M, although higher K.sub.D values may be tolerated where the k.sub.off is sufficiently low or the k.sub.on is sufficiently high.

[0099] In certain embodiments, the single domain polypeptide comprises an immunoglobulin (Ig) variable domain. The Ig variable domain may, for example, be selected from the group consisting of: a human V.sub.L domain, a human V.sub.H domain and a camelid V.sub.HH domain. One, two, three or more loops of the Ig variable domain may participate in binding to VEGFR-2, and typically any of the loops known as CDR1, CDR2 or CDR3 will participate in VEGFR-2 binding.

[0100] In certain embodiments, the single domain polypeptide comprises an immunoglobulin-like domain. One, two, three or more loops of the immunoglobulin-like domain may participate in binding to VEGFR-2. A preferred immunoglobulin-like domain is a fibronectin type III (Fn3) domain. Such domain may comprise, in order from N-terminus to C-terminus, a beta or beta-like strand, A; a loop, AB; a beta strand, B; a loop, BC; a beta strand C; a loop CD; a beta strand D; a loop DE; a beta strand F; a loop FG; and a beta or beta-like strand G.

[0101] Optionally, any or all of loops AB, BC, CD, DE, EF and FG may participate in VEGFR-2 binding, although preferred loops are BC, DE and FG. A preferred Fn3 domain is an Fn3 domain derived from human fibronectin, particularly the 10.sup.th Fn3 domain of fibronectin, referred to as .sup.10Fn3. It should be noted that none of VEGFR-2 binding polypeptides disclosed herein have an amino acid sequence that is identical to native .sup.10Fn3; the sequence has been modified to obtain VEGFR-2 binding proteins, but proteins having the basic structural features of .sup.10Fn3, and particularly those retaining recognizable sequence homology to the native .sup.10Fn3 are nonetheless referred to herein as ".sup.10Fn3 polypeptides". This nomenclature is similar to that found in the antibody field where, for example, a recombinant antibody V.sub.L domain generated against a particular target protein may not be identical to any naturally occurring V.sub.L domain but nonetheless the protein is recognizably a V.sub.L protein. A .sup.10Fn3 polypeptide may be at least 60%, 65%, 70%, 75%, 80%, 85%, or 90% identical to the human .sup.10Fn3 domain, shown in SEQ ID NO:5. Much of the variability will generally occur in one or more of the loops. Each of the beta or beta-like strands of a .sup.10Fn3 polypeptide may consist essentially of an amino acid sequence that is at least 80%, 85%, 90%, 95% or 100% identical to the sequence of a corresponding beta or beta-like strand of SEQ ID NO: 5, provided that such variation does not disrupt the stability of the polypeptide in physiological conditions. A .sup.10Fn3 polypeptide may have a sequence in each of the loops AB, CD, and EF that consists essentially of an amino acid sequence that is at least 80%, 85%, 90%, 95% or 100% identical to the sequence of a corresponding loop of SEQ ID NO:5. In many instances, any or all of loops BC, DE, and FG will be poorly conserved relative to SEQ ID NO:5. For example, all of loops BC, DE, and FG may be less than 20%, 10%, or 0% identical to their corresponding loops in SEQ ID NO:5.

[0102] In certain embodiments, the disclosure provides a non-antibody polypeptide comprising a domain having an immunoglobulin-like fold that binds to VEGFR-2. The non-antibody polypeptide may have a molecular weight of less than 20 kDa, or less than 15 kDa and will generally be derived (by, for example, alteration of the amino acid sequence) from a reference, or "scaffold", protein, such as an Fn3 scaffold. The non-antibody polypeptide may bind VEGFR-2 with a K.sub.D less than 10.sup.-6M, or less than 10.sup.-7M, less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M. The unaltered reference protein either will not meaningfully bind to VEGFR-2 or will bind with a K.sub.D of greater than 10.sup.-6M. The non-antibody polypeptide may inhibit VEGFR-2 signaling, particularly where the non-antibody polypeptide has a K.sub.D of less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M, although higher K.sub.D values may be tolerated where the k.sub.off is sufficiently low (e.g., less than 5.times.10.sup.-4s.sup.-1). The immunoglobulin-like fold may be a .sup.10Fn3 polypeptide.

[0103] In certain embodiments, the disclosure provides a polypeptide comprising a single domain having an immunoglobulin fold that binds to VEGFR-2. The polypeptide may have a molecular weight of less than 20 kDa, or less than 15 kDa and will generally be derived (by, for example, alteration of the amino acid sequence) from a variable domain of an immunoglobulin. The polypeptide may bind VEGFR-2 with a K.sub.D less than 10.sup.-6 M, or less than 10.sup.-7M, less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M. The polypeptide may inhibit VEGFR-2 signaling, particularly where the polypeptide has a K.sub.D of less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M, although higher K.sub.D values may be tolerated where the k.sub.off is sufficiently low or where the k.sub.on is sufficiently high. In some embodiments, the polypeptide comprises an amino acid sequence that is at least 80% identical to SEQ NO: 5. In some embodiments, the polypeptide comprises an amino acid sequence selected from the group consisting of any of SEQ ID NOs: 6-183, 186-197, 199 and 241-310. In some embodiments, the polypeptide further comprises PEG.

[0104] In certain aspects, the disclosure provides sustained-release delivery systems that deliver short peptide sequences that mediate VEGFR-2 binding. Such sequences may mediate VEGFR-2 binding in an isolated form or when inserted into a particular protein structure, such as an immunoglobulin or immunoglobulin-like domain. Examples of such sequences include those disclosed (such as SEQ ID NOs: 6-183, 186-197, 199 and 241-310) and other sequences that are at least 85%, 90%, or 95% identical to SEQ ID NO:5 to such sequences and retain VEGFR-2 binding activity. Accordingly, the disclosure provides substantially pure polypeptides comprising an amino acid sequence that is at least 85% identical to the sequence of any of such sequences, wherein said polypeptide binds to a VEGFR-2 and competes with an VEGF species for binding to VEGFR-2. Examples of such polypeptides include a polypeptide comprising an amino acid sequence that is at least 80%, 85%, 90%, 95% or 100% identical to an amino acid sequence of SEQ ID: 6-183, 186-197, 199 and 241-310. Preferably such polypeptides will inhibit a biological activity of a VEGF and may bind to VEGFR-2 with a K.sub.D less than 10.sup.-6 M, or less than 10.sup.-7M, less than 5.times.10.sup.-8M, less than 10.sup.-8M or less than 10.sup.-9M.

[0105] In certain embodiments, any of the VEGFR-2 binding polypeptides described herein may be bound to one or more additional moieties, including, for example, a moiety that also binds to VEGFR-2 (e.g., a second identical or different VEGFR-2 binding polypeptide), a moiety that binds to a different target (e.g., to create a dual-specificity binding agent), a labeling moiety, a moiety that facilitates protein purification or a moiety that provides improved pharmacokinetics. Improved pharmacokinetics may be assessed according to the perceived therapeutic need. Often it is desirable to increase bioavailability and/or increase the time between doses, possibly by increasing the time that a protein remains available in the serum after dosing. In some instances, it is desirable to improve the continuity of the serum concentration of the protein over time (e.g., decrease the difference in serum concentration of the protein shortly after administration and shortly before the next administration). Moieties that tend to slow clearance of a protein from the blood include polyethylene glycol, sugars (e.g. sialic acid), and well-tolerated protein moieties (e.g., Fc fragment or serum albumin). The single domain polypeptide may be attached to a moiety that reduces the clearance rate of the polypeptide in a mammal (e.g., mouse, rat, or human) by greater than three-fold relative to the unmodified polypeptide. Other measures of improved pharmacokinetics may include serum half-life, which is often divided into an alpha phase and a beta phase. Either or both phases may be improved significantly by addition of an appropriate moiety. Where polyethylene glycol is employed, one or more PEG molecules may be attached at different positions in the protein, and such attachment may be achieved by reaction with amines, thiols or other suitable reactive groups. Pegylation may be achieved by site-directed pegylation, wherein a suitable reactive group is introduced into the protein to create a site where pegylation preferentially occurs. In a preferred embodiment, the protein is modified so as to have a cysteine residue at a desired position, permitting site directed pegylation on the cysteine. PEG may vary widely in molecular weight and may be branched or linear. Notably, the present disclosure establishes that pegylation is compatible with target binding activity of .sup.10Fn3 polypeptides and, further, that pegylation does improve the pharmacokinetics of such polypeptides. Accordingly, in one embodiment, the disclosure provides pegylated forms of .sup.10Fn3 polypeptides, regardless of the target that can be bound by such polypeptides.

Nucleic Acids and Production of Polypeptides

[0106] Polypeptides of the present invention can be produced using any standard methods known in the art. In one example, the polypeptides are produced by recombinant DNA methods by inserting a nucleic acid sequence (e.g., a cDNA) encoding the polypeptide into a recombinant expression vector and expressing the DNA sequence under conditions promoting expression.

[0107] Nucleic acids encoding any of the various polypeptides disclosed herein may be synthesized chemically. Codon usage may be selected so as to improve expression in a cell. Such codon usage will depend on the cell type selected. Specialized codon usage patterns have been developed for E. coli and other bacteria, as well as mammalian cells, plant cells, yeast cells and insect cells. See for example: Mayfield et al., Proc Natl Acad Sci USA. 2003 Jan. 21; 100(2):438-42; Sinclair et al. Protein Expr Purif. 2002 October; 26(1):96-105; Connell N D. Curr Opin Biotechnol. 2001 October; 12(5):446-9; Makrides et al. Microbiol. Rev. 1996 September; 60(3):512-38; and Sharp et al. Yeast. 1991 October; 7(7):657-78.

[0108] Examples of nucleic acid sequences encoding a CT-01 polypeptide disclosed herein are:

TABLE-US-00009 SEQ ID NO:184 atgggcgaagttgttgctgcgacccccaccagcctactgatcagctggcg ccacccgcacttcccgactagatattacaggatcacttacggagaaacag gaggaaatagccctgtccaggagttcactgtgcctctgcagccccccaca gctaccatcagcggccttaaacctggagttgattataccatcactgtgta tgctgtcactgacggccggaacgggcgcctcctgagcatcccaatttcca ttaattaccgcacagaaattgacaaaccatgccag SEQ ID NO:185 atgggcgaagttgttgctgcgacccccaccagcctactgatcagctggcg ccacccgcacttcccgactagatattacaggatcacttacggagaaacag gaggaaatagccctgtccaggagttcactgtgcctctgcagccccccaca gctaccatcagcggccttaaacctggagttgattataccatcactgtgta tgctgtcactgacggccggaacgggcgcctcctgagcatcccaatttcca ttaattaccgcaca

[0109] General techniques for nucleic acid manipulation are described for example in Sambrook et al., Molecular Cloning: A Laboratory Manual, Vols. 1-3, Cold Spring Harbor Laboratory Press, 2 ed., 1989, or F. Ausubel et al., Current Protocols in Molecular Biology (Green Publishing and Wiley-Interscience: New York, 1987) and periodic updates, herein incorporated by reference. The DNA encoding the polypeptide is operably linked to suitable transcriptional or translational regulatory elements derived from mammalian, viral, or insect genes. Such regulatory elements include a transcriptional promoter, an optional operator sequence to control transcription, a sequence encoding suitable mRNA ribosomal binding sites, and sequences that control the termination of transcription and translation. The ability to replicate in a host, usually conferred by an origin of replication, and a selection gene to facilitate recognition of transformants are additionally incorporated.

[0110] The recombinant DNA can also include any type of protein tag sequence that may be useful for purifying the protein. Examples of protein tags include but are not limited to a histidine tag, a FLAG tag, a myc tag, an HA tag, or a GST tag. Appropriate cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian cellular hosts can be found in Cloning Vectors: A Laboratory Manual, (Elsevier, New York, 1985), the relevant disclosure of which is hereby incorporated by reference.

[0111] The expression construct is introduced into the host cell using a method appropriate to the host cell, as will be apparent to one of skill in the art. A variety of methods for introducing nucleic acids into host cells are known in the art, including, but not limited to, electroporation; transfection employing calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran, or other substances; microprojectile bombardment; lipofection; and infection (where the vector is an infectious agent).

[0112] Suitable host cells include prokaryotes, yeast, mammalian cells, or bacterial cells. Suitable bacteria include gram negative or gram positive organisms, for example, E. coli or Bacillus spp. Yeast, preferably from the Saccharomyces species, such as S. cerevisiae, may also be used for production of polypeptides. Various mammalian or insect cell culture systems can also be employed to express recombinant proteins. Baculovirus systems for production of heterologous proteins in insect cells are reviewed by Luckow and Summers, (Bio/Technology, 6:47, 1988). Examples of suitable mammalian host cell lines include endothelial cells, COS-7 monkey kidney cells, CV-1, L cells, C127, 3T3, Chinese hamster ovary (CHO), human embryonic kidney cells, HeLa, 293, 293T, and BHK cell lines. Purified polypeptides are prepared by culturing suitable host/vector systems to express the recombinant proteins. For many applications, the small size of many of the polypeptides disclosed herein would make expression in E. coli as the preferred method for expression. The protein is then purified from culture media or cell extracts.

[0113] Proteins disclosed herein can also be produced using cell-translation systems. For such purposes the nucleic acids encoding the polypeptide must be modified to allow in vitro transcription to produce mRNA and to allow cell-free translation of the mRNA in the particular cell-free system being utilized (eukaryotic such as a mammalian or yeast cell-free translation system or prokaryotic such as a bacterial cell-free translation system.

[0114] VEGFR-binding polypeptides can also be produced by chemical synthesis (e.g., by the methods described in Solid Phase Peptide Synthesis, 2nd ed., 1984, The Pierce Chemical Co., Rockford, Ill.). Modifications to the protein can also be produced by chemical synthesis.

[0115] The polypeptide of the present invention can be purified by isolation/purification methods for proteins generally known in the field of protein chemistry. Non-limiting examples include extraction, recrystallization, salting out (e.g., with ammonium sulfate or sodium sulfate), centrifugation, dialysis, ultrafiltration, adsorption chromatography, ion exchange chromatography, hydrophobic chromatography, normal phase chromatography, reversed-phase chromatography, gel filtration, gel permeation chromatography, affinity chromatography, electrophoresis, countercurrent distribution or any combinations of these. After purification, polypeptides may be exchanged into different buffers and/or concentrated by any of a variety of methods known to the art, including, but not limited to, filtration and dialysis.

[0116] The purified polypeptide is preferably at least 85% pure, more preferably at least 95% pure, and most preferably at least 98% pure. Regardless of the exact numerical value of the purity, the polypeptide is sufficiently pure for use as a pharmaceutical product. The polypeptide is in particular free of endotoxins

Post-Translational Modifications of Polypeptides

[0117] In certain embodiments, the binding polypeptides of the invention may further comprise post-translational modifications. Exemplary post-translational protein modification include phosphorylation, acetylation, methylation, ADP-ribosylation, ubiquitination, glycosylation, carbonylation, sumoylation, biotinylation or addition of a polypeptide side chain or of a hydrophobic group. As a result, the modified soluble polypeptides may contain non-amino acid elements, such as lipids, poly- or mono-saccharide, and phosphates. A preferred form of glycosylation is sialylation, which conjugates one or more sialic acid moieties to the polypeptide. Sialic acid moieties improve solubility and serum half-life while also reducing the possible immunogeneticity of the protein. See, e.g., Raju et al. Biochemistry. 2001 Jul. 31; 40(30):8868-76. Effects of such non-amino acid elements on the functionality of a polypeptide may be tested for its antagonizing role in VEGFR-2 or VEGF function, e.g., its inhibitory effect on angiogenesis or on tumor growth.

[0118] In one specific embodiment of the present invention, modified forms of the subject soluble polypeptides comprise linking the subject soluble polypeptides to nonproteinaceous polymers. In one specific embodiment, the polymer is polyethylene glycol ("PEG"), polypropylene glycol, or polyoxyalkylenes, in the manner as set forth in U.S. Pat. No. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337. Examples of the modified polypeptide of the invention include PEGylated CT-322.

[0119] PEG is a well-known, water soluble polymer that is commercially available or can be prepared by ring-opening polymerization of ethylene glycol according to methods well known in the art (Sandler and Karo, Polymer Synthesis, Academic Press, New York, Vol. 3, pages 138-161). The term "PEG" is used broadly to encompass any polyethylene glycol molecule, without regard to size or to modification at an end of the PEG, and can be represented by the formula:

X--O(CH.sub.2CH.sub.2O).sub.n-1CH.sub.2CH.sub.2OH (1),

where n is 20 to 2300 and X is H or a terminal modification, e.g., a C.sub.1-4 alkyl. In one embodiment, the PEG of the invention terminates on one end with hydroxy or methoxy, i.e., X is H or CH.sub.3 ("methoxy PEG"). A PEG can contain further chemical groups which are necessary for binding reactions; which results from the chemical synthesis of the molecule; or which is a spacer for optimal distance of parts of the molecule. In addition, such a PEG can consist of one or more PEG side-chains which are linked together. PEGs with more than one PEG chain are called multiarmed or branched PEGs. Branched PEGs can be prepared, for example, by the addition of polyethylene oxide to various polyols, including glycerol, pentaerythriol, and sorbitol. For example, a four-armed branched PEG can be prepared from pentaerythriol and ethylene oxide. Branched PEG are described in, for example, EP-A 0 473 084 and U.S. Pat. No. 5,932,462. One form of PEGs includes two PEG side-chains (PEG2) linked via the primary amino groups of a lysine (Monfardini, C., et al., Bioconjugate Chem. 6 (1995) 62-69).

[0120] In a preferred embodiment, the pegylated .sup.10Fn3 polypeptide is produced by site-directed pegylation, particularly by conjugation of PEG to a cysteine moiety at the N- or C-terminus. Accordingly, the present disclosure provides a target-binding .sup.10Fn3 polypeptide with improved pharmacokinetic properties, the polypeptide comprising: a .sup.10Fn3 domain having from about 80 to about 150 amino acids, wherein at least one of the loops of said .sup.10Fn3 domain participate in target binding; and a covalently bound PEG moiety, wherein said .sup.10Fn3 polypeptide binds to the target with a K.sub.D of less than 100 nM and has a clearance rate of less than 30 mL/hr/kg in a mammal. The PEG moiety may be attached to the .sup.10Fn3 polypeptide by site directed pegylation, such as by attachment to a Cys residue, where the Cys residue may be positioned at the N-terminus of the .sup.10Fn3 polypeptide or between the N-terminus and the most N-terminal beta or beta-like strand or at the C-terminus of the .sup.10Fn3 polypeptide or between the C-terminus and the most C-terminal beta or beta-like strand. A Cys residue may be situated at other positions as well, particularly any of the loops that do not participate in target binding. A PEG moiety may also be attached by other chemistry, including by conjugation to amines.

[0121] PEG conjugation to peptides or proteins generally involves the activation of PEG and coupling of the activated PEG-intermediates directly to target proteins/peptides or to a linker, which is subsequently activated and coupled to target proteins/peptides (see Abuchowski, A. et al, J. Biol. Chem., 252, 3571 (1977) and J. Biol. Chem., 252, 3582 (1977), Zalipsky, et al., and Harris et. al., in: Poly(ethylene glycol) Chemistry: Biotechnical and Biomedical Applications; (J. M. Harris ed.) Plenum Press: New York, 1992; Chap. 21 and 22). It is noted that a binding polypeptide containing a PEG molecule is also known as a conjugated protein, whereas the protein lacking an attached PEG molecule can be referred to as unconjugated.

[0122] A variety of molecular mass forms of PEG can be selected, e.g., from about 1,000 Daltons (Da) to 100,000 Da (n is 20 to 2300), for conjugating to VEGFR-2 binding polypeptides. The number of repeating units "n" in the PEG is approximated for the molecular mass described in Daltons. It is preferred that the combined molecular mass of PEG on an activated linker is suitable for pharmaceutical use. Thus, in one embodiment, the molecular mass of the PEG molecules does not exceed 100,000 Da. For example, if three PEG molecules are attached to a linker, where each PEG molecule has the same molecular mass of 12,000 Da (each n is about 270), then the total molecular mass of PEG on the linker is about 36,000 Da (total n is about 820). The molecular masses of the PEG attached to the linker can also be different, e.g., of three molecules on a linker two PEG molecules can be 5,000 Da each (each n is about 110) and one PEG molecule can be 12,000 Da (n is about 270).

[0123] In a specific embodiment of the invention, a VEGFR-2 binding polypeptide is covalently linked to one poly(ethylene glycol) group of the formula: --CO--(CH.sub.2).sub.x--(OCH.sub.2CH.sub.2).sub.m--OR, with the --CO (i.e. carbonyl) of the poly(ethylene glycol) group forming an amide bond with one of the amino groups of the binding polypeptide; R being lower alkyl; x being 2 or 3; m being from about 450 to about 950; and n and m being chosen so that the molecular weight of the conjugate minus the binding polypeptide is from about 10 to 40 kDa. In one embodiment, an binding polypeptide's .epsilon.-amino group of a lysine is the available (free) amino group.

[0124] The above conjugates may be more specifically presented by formula (II): P--NHCO--(CH.sub.2).sub.x--(OCH.sub.2CH.sub.2).sub.m--OR (II), wherein P is the group of a binding polypeptide as described herein, (i.e. without the amino group or amino groups which form an amide linkage with the carbonyl shown in formula (II); and wherein R is lower alkyl; x is 2 or 3; m is from about 450 to about 950 and is chosen so that the molecular weight of the conjugate minus the binding polypeptide is from about 10 to about 40 kDa. As used herein, the given ranges of "m" have an orientational meaning. The ranges of "m" are determined in any case, and exactly, by the molecular weight of the PEG group.

[0125] One skilled in the art can select a suitable molecular mass for PEG, e.g., based on how the pegylated binding polypeptide will be used therapeutically, the desired dosage, circulation time, resistance to proteolysis, immunogenicity, and other considerations. For a discussion of PEG and its use to enhance the properties of proteins, see N. V. Katre, Advanced Drug Delivery Reviews 10: 91-114 (1993).

[0126] In one embodiment of the invention, PEG molecules may be activated to react with amino groups on a binding polypeptide, such as with lysines (Bencham C. O. et al., Anal. Biochem., 131, 25 (1983); Veronese, F. M. et al., Appl. Biochem., 11, 141 (1985).; Zalipsky, S. et al., Polymeric Drugs and Drug Delivery Systems, adrs 9-110 ACS Symposium Series 469 (1999); Zalipsky, S. et al., Europ. Polym. J., 19, 1177-1183 (1983); Delgado, C. et al., Biotechnology and Applied Biochemistry, 12, 119-128 (1990)).

[0127] In one specific embodiment, carbonate esters of PEG are used to form the PEG-binding polypeptide conjugates. N,N'-disuccinimidylcarbonate (DSC) may be used in the reaction with PEG to form active mixed PEG-succinimidyl carbonate that may be subsequently reacted with a nucleophilic group of a linker or an amino group of a binding polypeptide (see U.S. Pat. No. 5,281,698 and U.S. Pat. No. 5,932,462). In a similar type of reaction, 1,1'-(dibenzotriazolyl)carbonate and di-(2-pyridyl)carbonate may be reacted with PEG to form PEG-benzotriazolyl and PEG-pyridyl mixed carbonate (U.S. Pat. No. 5,382,657), respectively.

[0128] Pegylation of a .sup.10Fn3 polypeptide can be performed according to the methods of the state of the art, for example by reaction of the binding polypeptide with electrophilically active PEGs (supplier: Shearwater Corp., USA, www.shearwatercorp.com). Preferred PEG reagents of the present invention are, e.g., N-hydroxysuccinimidyl propionates (PEG-SPA), butanoates (PEG-SBA), PEG-succinimidyl propionate or branched N-hydroxysuccinimides such as mPEG2--NHS (Monfardini, C., et al., Bioconjugate Chem. 6 (1995) 62-69). Such methods may used to pegylated at an E-amino group of a binding polypeptide lysine or the N-terminal amino group of the binding polypeptide.

[0129] In another embodiment, PEG molecules may be coupled to sulfhydryl groups on a binding polypeptide (Sartore, L., et al., Appl. Biochem. Biotechnol., 27, 45 (1991); Morpurgo et al., Biocon. Chem., 7, 363-368 (1996); Goodson et al., Bio/Technology (1990) 8, 343; U.S. Pat. No. 5,766,897). U.S. Pat. Nos. 6,610,281 and 5,766,897 describes exemplary reactive PEG species that may be coupled to sulfhydryl groups.

[0130] In some embodiments where PEG molecules are conjugated to cysteine residues on a binding polypeptide, the cysteine residues are native to the binding polypeptide, whereas in other embodiments, one or more cysteine residues are engineered into the binding polypeptide. Mutations may be introduced into an binding polypeptide coding sequence to generate cysteine residues. This might be achieved, for example, by mutating one or more amino acid residues to cysteine. Preferred amino acids for mutating to a cysteine residue include serine, threonine, alanine and other hydrophilic residues. Preferably, the residue to be mutated to cysteine is a surface-exposed residue. Algorithms are well-known in the art for predicting surface accessibility of residues based on primary sequence or a protein. Alternatively, surface residues may be predicted by comparing the amino acid sequences of binding polypeptides, given that the crystal structure of the framework based on which binding polypeptides are designed and evolved has been solved (see Himanen et al., Nature. (2001) 20-27; 414(6866):933-8) and thus the surface-exposed residues identified. In one embodiment, cysteine residues are introduced into binding polypeptides at or near the N- and/or C-terminus, or within loop regions.

[0131] In some embodiments, the pegylated binding polypeptide comprises a PEG molecule covalently attached to the alpha amino group of the N-terminal amino acid. Site specific N-terminal reductive amination is described in Pepinsky et al., (2001) JPET, 297, 1059, and U.S. Pat. No. 5,824,784. The use of a PEG-aldehyde for the reductive amination of a protein utilizing other available nucleophilic amino groups is described in U.S. Pat. No. 4,002,531, in Wieder et al., (1979) J. Biol. Chem. 254, 12579, and in Chamow et al., (1994) Bioconjugate Chem. 5, 133.

[0132] In another embodiment, pegylated binding polypeptide comprises one or more PEG molecules covalently attached to a linker, which in turn is attached to the alpha amino group of the amino acid residue at the N-terminus of the binding polypeptide. Such an approach is disclosed in U.S. Patent Publication No. 2002/0044921 and in WO94/01451.

[0133] In one embodiment, a binding polypeptide is pegylated at the C-terminus. In a specific embodiment, a protein is pegylated at the C-terminus by the introduction of C-terminal azido-methionine and the subsequent conjugation of a methyl-PEG-triarylphosphine compound via the Staudinger reaction. This C-terminal conjugation method is described in Cazalis et al., C-Terminal Site-Specific PEGylation of a Truncated Thrombomodulin Mutant with Retention of Full Bioactivity, Bioconjug Chem. 2004; 15(5): 1005-1009.

[0134] Monopegylation of a binding polypeptide can also be produced according to the general methods described in WO 94/01451. WO 94/01451 describes a method for preparing a recombinant polypeptide with a modified terminal amino acid alpha-carbon reactive group. The steps of the method involve forming the recombinant polypeptide and protecting it with one or more biologically added protecting groups at the N-terminal alpha-amine and C-terminal alpha-carboxyl. The polypeptide can then be reacted with chemical protecting agents to selectively protect reactive side chain groups and thereby prevent side chain groups from being modified. The polypeptide is then cleaved with a cleavage reagent specific for the biological protecting group to form an unprotected terminal amino acid alpha-carbon reactive group. The unprotected terminal amino acid alpha-carbon reactive group is modified with a chemical modifying agent. The side chain protected terminally modified single copy polypeptide is then deprotected at the side chain groups to form a terminally modified recombinant single copy polypeptide. The number and sequence of steps in the method can be varied to achieve selective modification at the N- and/or C-terminal amino acid of the polypeptide.

[0135] The ratio of a binding polypeptide to activated PEG in the conjugation reaction can be from about 1:0.5 to 1:50, between from about 1:1 to 1:30, or from about 1:5 to 1:15. Various aqueous buffers can be used in the present method to catalyze the covalent addition of PEG to the binding polypeptide. In one embodiment, the pH of a buffer used is from about 7.0 to 9.0. In another embodiment, the pH is in a slightly basic range, e.g., from about 7.5 to 8.5. Buffers having a pKa close to neutral pH range may be used, e.g., phosphate buffer.

[0136] Conventional separation and purification techniques known in the art can be used to purify PEGylated binding polypeptide, such as size exclusion (e.g. gel filtration) and ion exchange chromatography. Products may also be separated using SDS-PAGE. Products that may be separated include mono-, di-, tri- poly- and un-pegylated binding polypeptide, as well as free PEG. The percentage of mono-PEG conjugates can be controlled by pooling broader fractions around the elution peak to increase the percentage of mono-PEG in the composition. About ninety percent mono-PEG conjugates represents a good balance of yield and activity. Compositions in which, for example, at least ninety-two percent or at least ninety-six percent of the conjugates are mono-PEG species may be desired. In an embodiment of this invention the percentage of mono-PEG conjugates is from ninety percent to ninety-six percent.

[0137] In one embodiment, PEGylated binding polypeptide of the invention contain one, two or more PEG moieties. In one embodiment, the PEG moiety(ies) are bound to an amino acid residue which is on the surface of the protein and/or away from the surface that contacts the target ligand. In one embodiment, the combined or total molecular mass of PEG in PEG-binding polypeptide is from about 3,000 Da to 60,000 Da, optionally from about 10,000 Da to 36,000 Da. In a one embodiment, the PEG in pegylated binding polypeptide is a substantially linear, straight-chain PEG.

[0138] In one embodiment of the invention, the PEG in pegylated binding polypeptide is not hydrolyzed from the pegylated amino acid residue using a hydroxylamine assay, e.g., 450 mM hydroxylamine (pH 6.5) over 8 to 16 hours at room temperature, and is thus stable. In one embodiment, greater than 80% of the composition is stable mono-PEG-binding polypeptide, more preferably at least 90%, and most preferably at least 95%.

[0139] In another embodiment, the pegylated binding polypeptides of the invention will preferably retain at least 25%, 50%, 60%, 70% least 80%, 85%, 90%, 95% or 100% of the biological activity associated with the unmodified protein. In one embodiment, biological activity refers to its ability to bind to VEGFR-2, as assessed by K.sub.D, k.sub.on, or k.sub.off. In one specific embodiment, the pegylated binding polypeptide protein shows an increase in binding to VEGFR relative to unpegylated binding polypeptide.

[0140] The serum clearance rate of PEG-modified polypeptide may be decreased by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or even 90%, relative to the clearance rate of the unmodified binding polypeptide. The PEG-modified polypeptide may have a half-life (t.sub.1/2) which is enhanced relative to the half-life of the unmodified protein. The half-life of PEG-binding polypeptide may be enhanced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 250%, 300%, 400% or 500%, or even by 1000% relative to the half-life of the unmodified binding polypeptide. In some embodiments, the protein half-life is determined in vitro, such as in a buffered saline solution or in serum. In other embodiments, the protein half-life is an in vivo half life, such as the half-life of the protein in the serum or other bodily fluid of an animal.

Therapeutic Formulations and Modes of Administration

[0141] The present invention provides sustained-release intraocular drug delivery systems that are useful, in particular, for inhibiting VEGF biological activity. Techniques and dosages for administration vary depending on the type of specific polypeptide and the specific condition being treated but can be readily determined by the skilled artisan. In general, regulatory agencies require that a protein reagent to be used as a therapeutic be formulated so as to have acceptably low levels of pyrogens. Accordingly, therapeutic formulations will generally be distinguished from other formulations in that they are substantially pyrogen free, or at least contain no more than acceptable levels of pyrogen as determined by the appropriate regulatory agency (e.g., FDA). A pyrogen may be an endotoxin or exotoxin. In some embodiments, the drug delivery system is substantially endotoxin free.

[0142] Therapeutic compositions of the present invention may be administered with a pharmaceutically acceptable diluent, carrier, or excipient, in unit dosage form. Methods well known in the art for making formulations are found, for example, in "Remington: The Science and Practice of Pharmacy" (20th ed., ed. A. R. Gennaro A R., 2000, Lippincott Williams & Wilkins, Philadelphia, Pa.). Formulations for parenteral administration may, for example, contain excipients, sterile water, saline, polyalkylene glycols such as polyethylene glycol, oils of vegetable origin, or hydrogenated napthalenes. Biocompatible, biodegradable lactide polymer, lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be used to control the release of the compounds. Nanoparticulate formulations (e.g., biodegradable nanoparticles, solid lipid nanoparticles, liposomes) may be used to control the biodistribution of the compounds. Other potentially useful parenteral delivery systems include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. The concentration of the compound in the formulation varies depending upon a number of factors, including the dosage of the drug to be administered, and the route of administration.

[0143] The antiangiogenic polypeptide may be optionally administered as a pharmaceutically acceptable salt, such as non-toxic acid addition salts or metal complexes that are commonly used in the pharmaceutical industry. Examples of acid addition salts include organic acids such as acetic, lactic, pamoic, maleic, citric, malic, ascorbic, succinic, benzoic, palmitic, suberic, salicylic, tartaric, methanesulfonic, toluenesulfonic, or trifluoroacetic acids or the like; polymeric acids such as tannic acid, carboxymethyl cellulose, or the like; and inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid phosphoric acid, or the like. Metal complexes include zinc, iron, and the like. In one example, the polypeptide is formulated in the presence of sodium acetate to increase thermal stability.

[0144] The antiangiogenic polypeptides may also be entrapped in microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsule and poly-(methylmethacylate) microcapsule, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).

[0145] The formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.

[0146] A therapeutically effective dose refers to a dose that produces the therapeutic effects for which it is administered. The exact dose will depend on the disorder to be treated, and may be ascertained by one skilled in the art using known techniques. In addition, as is known in the art, adjustments for age as well as the body weight, general health, sex, diet, time of administration, drug interaction, and the severity of the disease may be necessary, and will be ascertainable with routine experimentation by those skilled in the art.

[0147] In some embodiments, the sustained-release drug delivery system is a liquid or a gel composition, suitable for injection into the ocular region of a patient. In some embodiments, the sustained release drug delivery system is a biodegradable implant. The drug system is injected intraocularly, such as an intravitreal, subconjunctival injection, or subtenon injection; and the resulting implant releases drug over a predetermined interval of time. Typically, the implant biodegrades at the same rate that the drug is released; therefore, the injection site essentially resolves in time for the next injection.

[0148] Sustained-release drug delivery systems also include semipermeable matrices of solid hydrophobic polymers containing the proteins of the invention, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and y ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT.TM. (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated proteins of the invention may remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37.degree. C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S--S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.

[0149] In some embodiments, the sustained release drug delivery system utilizes the Atrigel.TM. system comprising lactide/glycolide copolymers as described in U.S. Patent Application 20060210604. In some embodiments, the sustained release drug delivery system utilizes a biodegradable PLGA intravitreal implant as described in U.S. Patent Application 20050244469.

[0150] Other drug delivery systems have been previously described and may be used to deliver the antiangiogenic polypeptide component. The following is a list of suitable implants that may be used in the drug delivery system of the invention. U.S. Pat. No. 5,501,856 discloses controlled release pharmaceutical preparations for intraocular implants to be applied to the interior of the eye after a surgical operation for disorders in retina/vitreous body or for glaucoma. U.S. Pat. No. 5,869,079 discloses combinations of hydrophilic and hydrophobic entities in a biodegradable sustained release implant, and describes a polylactic acid polyglycolic acid (PLGA) copolymer implant comprising dexamethasone. As shown by in vitro testing of the drug release kinetics, the 100-120 .mu.g 50/50 PLGA/dexamethasone implant disclosed did not show appreciable drug release until the beginning of the fourth week, unless a release enhancer, such as HPMC was added to the formulation. U.S. Pat. No. 5,824,072 discloses implants for introduction into a suprachoroidal space or an avascular region of the eye, and describes a methylcellulose (i.e. non-biodegradable) implant comprising dexamethasone. WO 9513765 discloses implants comprising active agents for introduction into a suprachoroidal or an avascular region of an eye for therapeutic purposes. U.S. Pat. Nos. 4,997,652 and 5,164,188 disclose biodegradable ocular implants comprising microencapsulated drugs, and describes implanting microcapsules comprising hydrocortisone succinate into the posterior segment of the eye. U.S. Pat. No. 5,164,188 discloses encapsulated agents for introduction into the suprachoroid of the eye, and describes placing microcapsules and plaques comprising hydrocortisone into the pars plana. U.S. Pat. Nos. 5,443,505 and 5,766,242 disclose implants comprising active agents for introduction into a suprachoroidal space or an avascular region of the eye, and describes placing microcapsules and plaques comprising hydrocortisone into the pars plana. Zhou et al. disclose a multiple-drug implant comprising 5-fluorouridine, triamcinolone, and human recombinant tissue plasminogen activator for intraocular management of proliferative vitreoretinopathy (PVR). Zhou, T, et al. (1998). Development of a multiple-drug delivery implant for intraocular management of proliferative vitreoretinopathy, Journal of Controlled Release 55: 281-295. U.S. Pat. No. 6,369,116 discusses an implant with a release modifier inserted within a scleral flap. EP 0 654256 discusses use of a scleral plug after surgery on a vitreous body, for plugging an incision. U.S. Pat. No. 4,863,457 discusses the use of a bioerodible implant to prevent failure of glaucoma filtration surgery by positioning the implant either in the subconjunctival region between the conjunctival membrane overlying it and the sclera beneath it or within the sclera itself within a partial thickness sclera flap. EP 488 401 discusses intraocular implants, made of certain polylactic acids, to be applied to the interior of the eye after a surgical operation for disorders of the retina/vitreous body or for glaucoma. EP 430539 discusses use of a bioerodible implant which is inserted in the suprachoroid.

[0151] The amount of drug delivery system administered will typically depend upon the desired properties of the biodegradable implant For example, the amount of drug delivery system can influence the length of time in which the antiangiogenic polypeptide component is released from the biodegradable implant Additionally, the amount of drug delivery system administered will typically depend upon the specific intended use (e.g., nature and stage/progression of the disease or disorder).

[0152] Specifically, the drug delivery system can be formulated to provide an implant that releases therapeutically effective amounts of an antiangiogenic polypeptide for at least one week, two weeks, one month, two months, three months, four months, five months, six months, nine months, twelve months or more. Specifically, the drug delivery system can be formulated for administration less than about once per day. More specifically, the drug delivery system can be formulated for administration less than about once per week, less than about once per month, more than about once per year, about once per week to about once per year, or about once per month to about once per year.

[0153] In some embodiments, less than 5 ml, 4 ml, 3 ml, 2 ml, 1 ml, 0.1 ml, 0.01 ml, or 0.001 ml is administered. Specifically, the drug delivery system administered can range from about 0.01 mL to about 10.0 mL, about 0.05 mL to about 1.5 mL, about 0.1 mL to about 1.0 mL, or about 0.2 mL to about 0.8 mL.

[0154] The antiangiogenic polypeptide component can be present in any effective, suitable and appropriate amount. For example, polypeptide component can be present up to about 70 wt. % of the drug delivery system, up to about 60 wt. % of the drug delivery system, up to about 40 wt. % of the drug delivery system, up to about 20 wt. % of the drug delivery system, 10 wt. % of the drug delivery system, up to about 5 wt. % of the drug delivery system, up to about 1 wt. % of the drug delivery system, or up to about 0.1 wt. % of the drug delivery system.

[0155] The drug delivery system will effectively deliver the antiangiogenic polypeptide component to mammalian tissue at a suitable, effective, safe, and appropriate dosage. For example, the drug delivery system can effectively deliver the antiangiogenic polypeptide component to mammalian tissue at a dosage of more than about 0.001 picogram/kilogram/day, more than about 0.01 picogram/kilogram/day, more than about 0.1 picogram/kilogram/day, or more than about 1 picogram/kilogram/day. Alternatively, the drug delivery system can effectively deliver the antiangiogenic polypeptide component to mammalian tissue at a dosage of up to about 100 milligram/kilogram/day, up to about 50 milligram/kilogram/day, up to about 10 milligram/kilogram/day, or up to about 1 milligram/kilogram/day.

[0156] More specifically, the drug delivery system can effectively deliver the antiangiogenic polypeptide component to mammalian tissue at a dosage of about 0.001 picogram/kilogram/day to about 100 milligram/kilogram/day; about 0.01 picogram/kilogram/day to about 50 milligram/kilogram/day; about 0.1 picogram/kilogram/day to about 10 milligram/kilogram/day; or about 1 picogram/kilogram/day to about 1 milligram/kilogram/day.

[0157] The sustained-release intraocular drug delivery system can further comprise analgesics, anesthetics, anti-infective agents, or anti-steroidal agents. Suitable analgesics include, e.g., acetaminophen, phenylpropanolamine HCl, chlorpheniramine maleate, hydrocodone bitartrate, acetaminophen elixir, diphenhydramine HCl, pseudoephedrine HCl, dextromethorphan HBr, guaifenesin, doxylamine succinate, pamabron, clonidine hydrochloride, tramadol hydrochloride, carbamazepine, sodium hyaluronate, lidocaine, hylan, Arnica Montana, radix (mountain arnica), Calendula officinalis (marigold), Hamamelis (witch hazel), Millefolium (milfoil), Belladonna (deadly nightshade), Aconitum napellus (monkshood), Chamomilla (chamomile), Symphytum officinale (comfrey), Bellis perennis (daisy), Echinacea angustifolia (narrow-leafed cone flower), Hypericum perforatum (St. John's wort), Hepar sulphuris calcareum (calcium sulfide), buprenorphine hydrochloride, nalbuphine hydrochloride, pentazocine hydrochloride, acetylsalicylic acid, salicylic acid, naloxone hydrochloride, oral transmucosal fentanyl citrate, morphine sulfate, propoxyphene napsylate, propoxyphene hydrochloride, meperidine hydrochloride, hydromorphone hydrochloride, fentanyl transdermal system, levorphanol tartrate, promethazine HCl, oxymorphone hydrochloride, levomethadyl acetate hydrochloride, oxycodone HCl, oxycodone, codeine phosphate, isometheptene mucate, dichloralphenazone, butalbital, naproxen sodium, diclofenac sodium, misoprostol, diclofenac potassium, celecoxib, sulindac, oxaprozin, salsalate, diflunisal, naproxen, piroxicam, indomethacin, indomethacin sodium trihydrate, etodolac, meloxicam, ibuprofen, fenoprofen calcium, ketoprofen, mefenamic acid, nabumetone, tolmetin sodium, ketorolac tromethamine, choline magnesium trisalicylate, and rofecoxib.

[0158] Suitable anesthetics include: propofol, halothane, desflurane, midazolam HCl, epinephrine, levobupivacaine, etidocaine hydrochloride, ropivacaine HCl, chloroprocaine HCl, bupivacaine HCl, and lidocaine HCl.

[0159] Suitable anti-infective agents include, e.g., trimethoprim, sulfamethoxazole, clarithromycin, ganciclovir sodium, ganciclovir, daunorubicin citrate liposome, fluconazole, doxorubicin HCl liposome, foscamet sodium, interferon alfa-2b, atovaquone, rifabutun, trimetrexate glucoronate, itraconazole, ciclofovir, azithromycin, delavirdine mesylate, efavirenz, nevirapine, lamivudine/zidovudine, zalcitabine, didanosine, stavudine, abacavir sulfate, amprenavir, indinavir sulfate, saquinavir, saquinavir mesylate, ritonavir, nelfinavir, chloroquine hydrochloride, metronidazole, metronidazole hydrochloride, iodoquinol, albendazole, praziquantel, thiabendazole, ivermectin, mebendazole sulfate, tobramycin sulfate, tobramycin, azetreonam, cefotetan disodium, cefotetan, loracarbef, cefoxitin, meropenem, imipenemand cilastatin, cefazolin, cefaclor, ceftibuten, ceftizoxime, cefoperazone, cefuroxumeaxetil, cefprozil, ceftazidime, cefotaxime sodium, cefadroxil monohydrate, cephalexin, cephalexin hydrochloride, cefuroxime, cefazolin, cefamandole nafate, cefapime hydrochloride, cefdinir, ceftriaxone sodium, cefixme, cefpodoxime proxetil, dirithromycin, erythromycin, erythromycin ethylsuccinate, erythromycin stearate, erythromycin, sulfisoxazole acetyl, troleandomycin, azithromycin, clindamycin, clindamycin hydrochloride, colistimethate sodium, quinupristin/dalfopristin, vancomycin hydrochloride, amoxicillin, amoxicillin/calvulanate/potassium, penicillin G benzathine, penicillin G procaine, penicillin G potassium, carbenicillin indanyl sodium, piperacillin sodium, ticarcillin disodium, clavulanate potassium, ampicillin sodium/sulbactam sodium, tazobactam sodium, tetracycline HCl, demeclocycline hydrochloride, doxycycline hyclate, minocycline HCl, doxycycline monohydrate, oxytetracycline HCl, hydrocortisone acetate, doxycycline calcium, amphotericin B lipid, flucytosine, griseofulvin, terbinafine hydrochloride, ketoconazole, chloroquine hydrochloride, chloroquine phosphate, pyrimethamine, mefloquine hydrochloride, atovaquone and proguanil hydrochloride, hydroxychloroquine sulfate, ethambutol hydrochloride, aminosalicylic acid, rifapentine, rifampin, isoniazid, pyrazinamide, ethionamide, interferon alfa-n3, famciclovir, rimantadine hydrochloride, foscamet sodium, interferon alfacon-1, ribavirin, zanamivir, amantadine hydrochloride, palivizumab, oseltamivir phosphate, valacyclovir hydrochloride, nelfinavir mesylate, stavudine, acyclovir, acyclovir sodium, rifabutin, trimetrexate glucuronate, linezolid, moxifloxacin, moxifloxacin hydrochloride, ciprofloxacin, ciprofloxacin hydrochloride, ofloxacin, levofloxacin, lomefloxacin hydrochloride, nalidixic acid, norfloxacin, enoxacin, gatifloxacin, trovafloxacin mesylate, alatrofloxacin, sparfloxacin, aztreonam, nitrofurantoin monohydrate/macrocrystals, cefepime hydrochloride, fosfomycin tromethamine, neomycin sulfate-polymyxin B sulfate, imipenem, cilastatin, methenamine, methenamine mandelate, phenyl salicylate, atropine sulfate, hyoscyamine sulfate, benzoic acid, oxytetracycline hydrochloride, sulfamethizole, phenazopyridine hydrochloride, and sodium acid phosphate, monohydrate.

[0160] The steroidal anti-inflammatory agents that may be used in the ocular implants include, but are not limited to, 21-acetoxypregnenolone, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clobetasone, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort, flucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halometasone, halopredone acetate, hydrocortamate, hydrocortisone, loteprednol etabonate, mazipredone, medrysone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylamino-acetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, and any of their derivatives.

Exemplary Uses

[0161] The small size and stable structure of the disclosed polypeptides can be particularly valuable with respect to manufacturing of the drug, rapid clearance from the body for certain applications where rapid clearance is desired or formulation into novel delivery systems that are suitable or improved using a molecule with such characteristics.

[0162] On the basis of their efficacy as inhibitors of VEGF biological activity, the polypeptides of the invention are effective against a number of conditions associated with inappropriate angiogenesis, including but not limited to autoimmune disorders (e.g., rheumatoid arthritis, inflammatory bowel disease or psoriasis); cardiac disorders (e.g., atherosclerosis or blood vessel restenosis); retinopathies (e.g., proliferative retinopathies generally, diabetic retinopathy, age-related macular degeneration or neovascular glaucoma), renal disease (e.g., diabetic nephropathy, malignant nephrosclerosis, thrombotic microangiopathy syndromes; transplant rejection; inflammatory renal disease; glomerulonephritis; mesangioproliferative glomerulonephritis; haemolytic-uraemic syndrome; and hypertensive nephrosclerosis); hemangioblastoma; hemangiomas; thyroid hyperplasias; tissue transplantations; chronic inflammation; Meigs's syndrome; pericardial effusion; pleural effusion; autoimmune diseases; diabetes; endometriosis; chronic asthma; undesirable fibrosis (particularly hepatic fibrosis) and cancer, as well as complications arising from cancer, such as pleural effusion and ascites. Preferably, the VEGFR-binding polypeptides of the invention can be used for the treatment of prevention of hyperproliferative diseases or cancer and the metastatic spread of cancers. Non-limiting examples of cancers include bladder, blood, bone, brain, breast, cartilage, colon kidney, liver, lung, lymph node, nervous tissue, ovary, pancreatic, prostate, skeletal muscle, skin, spinal cord, spleen, stomach, testes, thymus, thyroid, trachea, urogenital tract, ureter, urethra, uterus, or vaginal cancer. Additional treatable conditions can be found in U.S. Pat. No. 6,524,583, herein incorporated by reference. Other references describing uses for VEGFR-2 binding polypeptides include: McLeod D S et al., Invest Opthalmol V is Sci. 2002 February; 43(2):474-82; Watanabe et al. Exp Dermatol. 2004 Nov.; 13(11):671-81; Yoshiji H et al., Gut. 2003 September; 52(9):1347-54; Verheul et al., Oncologist. 2000; 5 Suppl 1:45-50; Boldicke et al., Stem Cells. 2001; 19(1):24-36.

[0163] As described herein, angiogenesis-associated diseases include, but are not limited to, angiogenesis-dependent cancer, including, for example, solid tumors, blood born tumors such as leukemias, and tumor metastases; benign tumors, for example hemangiomas, acoustic neuromas, neurofibromas, trachomas, and pyogenic granulomas; inflammatory disorders such as immune and non-immune inflammation; chronic articular rheumatism and psoriasis; ocular angiogenic diseases, for example, diabetic retinopathy, retinopathy of prematurity, macular degeneration, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis; Osler-Webber Syndrome; myocardial angiogenesis; plaque neovascularization; telangiectasia; hemophiliac joints; angiofibroma; and wound granulation and wound healing; telangiectasia psoriasis scleroderma, pyogenic granuloma, cororany collaterals, ischemic limb angiogenesis, corneal diseases, rubeosis, arthritis, diabetic neovascularization, fractures, vasculogenesis, hematopoiesis.

[0164] In particular, the sustained-release intraocular drug delivery system is useful for the treatment of retinopathies, such as retinal vein occlusion, diabetic macular edema, diabetic retinopathy, retinopathy of prematurity, macular degeneration, age-related macular degeneration, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, and rubeosis.

[0165] In one embodiment, the drug delivery system administers a therapeutic component to ameliorate inflammation, and thus to control, reduce or prevent an inflammatory response or ameliorate the effects of an inflammatory response. In one embodiment, the therapeutic component is used to enhance reabsorption of inflammatory exudates. Decreasing the level of exudates in the eye reduces the inflammatory process and the ensuing hyperpermeable state that occurs with allergies, infection, responses to ocular photodynamic therapy (PDT) and laser treatments, after ocular surgery or trauma, etc.

[0166] In one embodiment, the therapeutic component is administered to ameliorate the scarring and adhesions that are a part of the inflammatory process. Adhesions are bands of scar tissue that bind two internal body surfaces. They are an inflammatory response to tissue damage, and occur as a normal part of any healing process. As one example, adhesions frequently occur during the post-surgical healing process during which tissues have experienced mechanical trauma. However, adverse effects can occur when internal surfaces bind, and adhesions may persist even after the original trauma has healed. Surgery to repair adhesions itself results in recurrent or additional adhesions. The presence of adhesions may also complicate surgical procedures, for example, ocular conjunctival adhesions may complicate subsequent glaucoma surgery.

[0167] Adhesions can occur following any type of trauma or surgery, including but not limited to ocular surgery. Examples of ocular surgery that may result in adhesions include glaucoma filtration operations (i.e., iridencleisis and trephination, pressure control valves), extraocular muscle surgery, diathermy or scleral buckling surgery for retinal detachment, and vitreous surgery. Examples of ocular trauma include penetrating ocular injuries, intraocular foreign body, procedures such as PDT, scatter laser threshold coagulation, refractive surgery, and blunt trauma.

[0168] In one embodiment, the therapeutic component ameliorates disorders with both a vascular proliferative component and a scarring component. As one example, the invention may be used in patients with the ocular disease pterygia. In these patients, fibrovascular proliferation results in scarring of the conjunctiva. An elevated, superficial, external ocular mass, termed a pterygium, forms and extends onto the corneal surface. Patients may experience symptoms of inflammation (e.g., redness, swelling, itching, irritation) and blurred vision. The mass itself may become inflamed, resulting in redness and ocular irritation. Left untreated, pterygia can distort the corneal topography, obscure the optical center of the cornea, and result in altered vision.

[0169] The process whereby scar tissue forms (scarring) can occur without new blood vessels being formed (neovascularization). However, the neovascularization process always results in scarring because of the cell proliferation that occurs with the formation of new vessels also results in the proliferation of fibroblasts, glial cells, etc. that result in scar tissue formation. The inventive method may be used to ameliorate the scarring process.

[0170] In one embodiment, the therapeutic component is administered to ameliorate inflammation of uveal tissues (uveitis, an inflammation of tissues in the middle layer of the eye, mainly the iris (iritis) and the ciliary body). Ocular inflammation may be associated with underlying systemic disease or autoimmunity, or may occur as a direct result of ocular trauma or infectious agents (bacterial, viral, fungal, etc.). Inflammatory reactions in adjacent tissues, e.g., keratitis, can induce a secondary uveitis. There are both acute and chronic forms of uveitis. The chronic form is frequently associated with many systemic disorders and most likely occurs due to immunopathological mechanisms.

[0171] Uveitis presents with ocular pain, photophobia and hyperlacrimation, with decreased visual acuity ranging from mild blur to significant vision loss. Hallmark signs of anterior uveitis are cells and flare in the anterior chamber. If the anterior chamber reaction is significant, small gray to brown endothelial deposits known as keratic precipitates may arise, leading to endothelial cell dysfunction and corneal edema. There may be adhesions to the lens capsule (posterior synechia) or the peripheral cornea (anterior synechia). Granulomatous nodules may appear on the surface of the iris stroma. Intraocular pressure is initially reduced due to secretory hypotony of the ciliary body but, as the reaction persists, inflammatory by-products may accumulate in the trabeculum. If this debris builds significantly, and if the ciliary body resumes its normal secretory output, the pressure may rise sharply, resulting in a secondary uveitic glaucoma.

[0172] A VEGFR-2 binding polypeptide can be administered alone or in combination with one or more additional therapies such as chemotherapy radiotherapy, immunotherapy, surgical intervention, or any combination of these. Long-term therapy is equally possible as is adjuvant therapy in the context of other treatment strategies, as described above.

[0173] In certain embodiments of such methods, one or more polypeptide therapeutic agents can be administered, together (simultaneously) or at different times (sequentially). In addition, polypeptide therapeutic agents can be administered with another type of compounds for treating cancer or for inhibiting angiogenesis.

[0174] In certain embodiments, the subject therapeutic agents of the invention can be used alone. Alternatively, the subject agents may be used in combination with conventional anti-cancer therapeutic approaches directed to treatment or prevention of proliferative disorders (e.g., tumor). For example, such methods can be used in prophylactic cancer prevention, prevention of cancer recurrence and metastases after surgery, and as an adjuvant of other conventional cancer therapy. The present invention recognizes that the effectiveness of conventional cancer therapies (e.g., chemotherapy, radiation therapy, phototherapy, immunotherapy, and surgery) can be enhanced through the use of a subject polypeptide therapeutic agent.

[0175] A wide array of conventional compounds have been shown to have anti-neoplastic activities. These compounds have been used as pharmaceutical agents in chemotherapy to shrink solid tumors, prevent metastases and further growth, or decrease the number of malignant cells in leukemic or bone marrow malignancies. Although chemotherapy has been effective in treating various types of malignancies, many anti-neoplastic compounds induce undesirable side effects. It has been shown that when two or more different treatments are combined, the treatments may work synergistically and allow reduction of dosage of each of the treatments, thereby reducing the detrimental side effects exerted by each compound at higher dosages. In other instances, malignancies that are refractory to a treatment may respond to a combination therapy of two or more different treatments.

[0176] When the drug delivery system of the present invention is administered in combination with a conventional anti-neoplastic agent, either concomitantly or sequentially, such drug delivery system may be found to enhance the therapeutic effect of the anti-neoplastic agent or overcome cellular resistance to such anti-neoplastic agent. This allows decrease of dosage of an anti-neoplastic agent, thereby reducing the undesirable side effects, or restores the effectiveness of an anti-neoplastic agent in resistant cells.

[0177] The therapeutic agents that can be combined with the sustained-release drug delivery system of the invention include diverse agents used in oncology practice (Reference: Cancer, Principles & Practice of Oncology, DeVita, V. T., Hellman, S., Rosenberg, S. A., 6th edition, Lippincott-Raven, Philadelphia, 2001), such as, merely to illustrate: abarelix, altretamine, aminoglutethimide, amsacrine, anastrozole, antide, asparaginase, AZD2171 (Recentin.TM.), Bacillus Calmette-Guerin/BCG (TheraCys.TM., TICE.TM.), bevacizumab (see U.S. Pat. No. 6,054,297; Avastin.TM.), bicalutamide, bleomycin, bortezomib (Velcade.TM.), buserelin, busulfan, campothecin, capecitabine, carboplatin, carmustine, cetuximab (Erbitux.TM.), chlorambucil, cisplatin, cladribine, clodronate, colchicine, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, dasatinib ((see U.S. Pat. No. 6,596,746 Sprycel.TM.), daunorubicin, dienestrol, diethylstilbestrol, dexamethasone, docetaxel (Taxotere.TM.), doxorubicin, Abx-EGF, epothilones, epirubicin, erlonitib (Tarceva.TM.), estradiol, estramustine, etoposide, exemestane, 5-fluorouracil, filgrastim, fludarabine, fludrocortisone, fluorouracil, fluoxymesterone, flutamide, fulvestrant, gefitinib (Iressa.TM.), gemcitabine (see U.S. Pat. No. 4,808,614; Gemzar.TM.), genistein, goserelin, hydroxyurea, idarubicin, ifosfamide, imatinib mesylate (see U.S. Pat. No. 5,521,184; Gleevac.TM.), interferon, irinotecan, ibritumomab (Zevalin.TM.), ironotecan, ixabepilone (BMS-247550), lapatinib (see U.S. Pat. No. 6,391,874; Tykreb.TM.), letrozole, leucovorin, leuprolide, levamisole, lomustine, mechlorethamine, medroxyprogesterone, megestrol, melphalan, mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, motesanib diphosphate (AMG 706) nilutamide, nocodazole, octreotide, oxaliplatin, paclitaxel (Taxol.TM.), pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rapamycin, rituximab (Rituxan.TM.), sorafenib (Nexavar.TM./Bayer BAY43-9006), streptozocin, suramin, sunitinib malate (see U.S. Pat. No. 6,573,293; Sutent.TM.), tamoxifen, temsirolimus (see U.S. Pat. No. 5,362,718; CCl-779), temozolomide (see U.S. Pat. No. 5,260,291; Temodar.TM.), teniposide, testosterone, thioguanine, thiotepa, titanocene dichloride, topotecan, toremifene, tositumomab (Bexxar.TM.), trastuzumab (U.S. Pat. No. 5,821,337; Herceptin.TM.), tretinoin, VEGF Trap (aflibercept; preparation described in U.S. Pat. No. 5,844,099), vinblastine, vincristine, vindesine, and vinorelbine, zoledronate.

[0178] Certain chemotherapeutic anti-tumor compounds may be categorized by their mechanism of action into, for example, following groups: anti-metabolites/anti-cancer agents, such as pyrimidine analogs (5-fluorouracil, floxuridine, capecitabine, gemcitabine and cytarabine) and purine analogs, folate antagonists and related inhibitors (mercaptopurine, thioguanine, pentostatin and 2-chlorodeoxyadenosine (cladribine)); antiproliferative/antimitotic agents including natural products such as vinca alkaloids (vinblastine, vincristine, and vinorelbine), microtubule disruptors such as taxane (paclitaxel, docetaxel), vincristin, vinblastin, nocodazole, epothilones and navelbine, epidipodophyllotoxins (etoposide, teniposide), DNA damaging agents (actinomycin, amsacrine, anthracyclines, bleomycin, busulfan, camptothecin, carboplatin, chlorambucil, cisplatin, cyclophosphamide, cytoxan, dactinomycin, daunorubicin, doxorubicin, epirubicin, hexamethylmelamineoxaliplatin, iphosphamide, melphalan, merchlorehtamine, mitomycin, mitoxantrone, nitrosourea, plicamycin, procarbazine, taxol, taxotere, teniposide, triethylenethiophosphoramide and etoposide (VP 16)); antibiotics such as dactinomycin (actinomycin D), daunorubicin, doxorubicin (adriamycin), idarubicin, anthracyclines, mitoxantrone, bleomycins, plicamycin (mithramycin) and mitomycin; enzymes (L-asparaginase which systemically metabolizes L-asparagine and deprives cells which do not have the capacity to synthesize their own asparagine); antiplatelet agents; antiproliferative/antimitotic alkylating agents such as nitrogen mustards (mechlorethamine, cyclophosphamide and analogs, melphalan, chlorambucil), ethylenimines and methylmelamines (hexamethylmelamine and thiotepa), alkyl sulfonates-busulfan, nitrosoureas (carmustine (BCNU) and analogs, streptozocin), trazenes--dacarbazinine (DTIC); antiproliferative/antimitotic antimetabolites such as folic acid analogs (methotrexate); platinum coordination complexes (cisplatin, carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide; hormones, hormone analogs (estrogen, tamoxifen, goserelin, bicalutamide, nilutamide) and aromatase inhibitors (letrozole, anastrozole); anticoagulants (heparin, synthetic heparin salts and other inhibitors of thrombin); fibrinolytic agents (such as tissue plasminogen activator, streptokinase and urokinase), aspirin, dipyridamole, ticlopidine, clopidogrel, abciximab; antimigratory agents; antisecretory agents (breveldin); immunosuppressives (cyclosporine, tacrolimus (FK-506), sirolimus (rapamycin), azathioprine, mycophenolate mofetil); anti-angiogenic compounds (TNP-470, genistein) and growth factor inhibitors (e.g., VEGF inhibitors, fibroblast growth factor (FGF) inhibitors); angiotensin receptor blocker; nitric oxide donors; anti-sense oligonucleotides; antibodies (trastuzumab); cell cycle inhibitors and differentiation inducers (tretinoin); mTOR inhibitors, topoisomerase inhibitors (doxorubicin (adriamycin), amsacrine, camptothecin, daunorubicin, dactinomycin, eniposide, epirubicin, etoposide, idarubicin and mitoxantrone, topotecan, irinotecan), corticosteroids (cortisone, dexamethasone, hydrocortisone, methylpednisolone, prednisone, and prenisolone); growth factor signal transduction kinase inhibitors; mitochondrial dysfunction inducers and caspase activators; and chromatin disruptors.

[0179] In certain embodiments, pharmaceutical compounds that may be used for combinatory anti-angiogenesis therapy include: (1) inhibitors of release of "angiogenic molecules," such as bFGF (basic fibroblast growth factor); (2) neutralizers of angiogenic molecules, such as an anti-.beta.bFGF antibodies; and (3) inhibitors of endothelial cell response to angiogenic stimuli, including collagenase inhibitor, basement membrane turnover inhibitors, angiostatic steroids, fungal-derived angiogenesis inhibitors, platelet factor 4, thrombospondin, arthritis drugs such as D-penicillamine and gold thiomalate, vitamin D.sub.3 analogs, alpha-interferon, and the like. For additional proposed inhibitors of angiogenesis, see Blood et al., Bioch. Biophys. Acta., 1032:89-118 (1990), Moses et al., Science, 248:1408-1410 (1990), Ingber et al., Lab. Invest., 59:44-51 (1988), and U.S. Pat. Nos. 5,092,885, 5,112,946, 5,192,744, 5,202,352, and 6573256. In addition, there are a wide variety of compounds that can be used to inhibit angiogenesis, for example, endostatin protein or derivatives, lysine binding fragments of angiostatin, melanin or melanin-promoting compounds, plasminogen fragments (e.g., Kringles 1-3 of plasminogen), tropoin subunits, antagonists of vitronectin .alpha..sub.v.beta..sub.3, peptides derived from Saposin B, antibiotics or analogs (e.g., tetracycline, or neomycin), dienogest-containing compositions, compounds comprising a MetAP-2 inhibitory core coupled to a peptide, the compound EM-138, chalcone and its analogs, and naaladase inhibitors. See, for example, U.S. Pat. Nos. 6,395,718, 6,462,075, 6,465,431, 6,475,784, 6,482,802, 6,482,810, 6,500,431, 6,500,924, 6,518,298, 6,521,439, 6,525,019, 6,538,103, 6,544,758, 6,544,947, 6,548,477, 6,559,126, and 6,569,845.

[0180] Depending on the nature of the combinatory therapy, administration of the drug delivery system of the invention may be continued while the other therapy is being administered and/or thereafter. Administration of the drug delivery system may be made in a single dose, or in multiple doses. In some instances, administration of the drug delivery system is commenced at least several days prior to the conventional therapy, while in other instances, administration is begun either immediately before or at the time of the administration of the conventional therapy.

EXAMPLES

[0181] The following examples are for the purposes of illustrating the invention, and should not be construed as limiting.

Example 1

Initial Identification of KDR Binding Molecules

[0182] A library of approximately 1013 RNA-protein fusion variants was constructed based on the scaffold of the tenth type 3 domain of human fibronectin with three randomized regions at positions 23-29, 52-55 and 77-86 (amino acid nos. are referenced to SEQ ID NO:5) (three loop library; Xu et al, Chemistry & Biology 9:933-942, 2002). Similar libraries were constructed containing randomized regions only at positions 23-29 and 77-86 (two loop library) or only at positions 77-86 (one loop library). A mixture of these three libraries was used for in vitro selection against the extracellular domain of human VEGFR-2 (KDR, extracellular domain, residues 1-764 fused to human IgG1 Fc). For the purposes of this application, the amino acid positions of the loops will be defined as residues 23-30 (BC Loop), 52-56 (DE Loop) and 77-87 (FG Loop). The target binding population was analyzed by DNA sequencing after six rounds of selection and was found to be diverse, with some replicates present. Proteins encoded by fifteen independent clones were screened for binding to KDR, (FIG. 1A) and the best binders were subsequently analyzed for inhibition of target binding in the presence of VEGF (FIG. 1B). Multiple clones were identified that inhibited KDR-VEGF binding, suggesting that these clones bound KDR at or near the natural ligand (VEGF) binding site. The ability of two of the binding molecules (VR28 and VR12) to directly inhibit VEGF-KDR interaction was evaluated in a BIAcore assay using immobilized VEGF and a mobile phase containing KDR-Fc with or without a selected binding protein. VR28 and, to a lesser extent, VR12, but not a non-competing clone (VR17), inhibited KDR binding to VEGF in a dose dependent manner (FIG. 1C). Finally, in addition to binding to purified recombinant KDR, VR28 also appeared to bind to KDR-expressing recombinant CHO cells, but not to control CHO cells (FIG. 1D).

[0183] The sequence of the binding loops of the VR28 clone is shown in the first row of Table 4. While VR28 was not the most abundant clone in the sequenced binding population (one copy out of 28 sequenced clone), its binding affinity to KDR was the best among the tested clones from this binding population, with a dissociation constant of 11-13 nM determined in a radioactive equilibrium binding assay (FIG. 3 and Table 5) and BIAcore assays (Table 7). There were no changes from wild type .sup.10Fn3 in the remaining scaffold portion of the molecule (following correction of an incidental scaffold change at position 69 that had no effect on binding). However, VR28 showed little inhibition of VEGF-KDR signaling in a VEGF-dependent cell proliferation assay. Thus, while the selection from the naive library yielded antibody mimics that interfered with the interaction between VEGF and KDR in biochemical binding studies, affinity improvements were useful for neutralizing function in a biological signal transduction assay.

Example 2

Affinity Maturation of Clone VR28

[0184] A mutagenesis strategy focusing on altering sequences only in the binding loops was employed. To initially test which loops were more likely to result in improvement, loop-directed hypermutagenic PCR was carried out to introduce up to 30% mutations independently into each loop of VR28. After three rounds of selection against KDR, multiple clones with improved binding to KDR-Fc were observed. Sequence analysis of the selection pools revealed that the majority of mutations were accumulated in the FG loop while the BC and DE loops remained almost intact. This result indicated that the FG loop was the most suitable target for further modification.

[0185] Consequently, a new library of approximately 1012 variants was constructed by altering the sequence of VR28 in the FG loop using oligonucleotide mutagenesis. For each of the FG loop positions (residues 77-86 [VAQNDHELIT (SEQ ID NO:198)] as well as the following Proline [residue 87]), a 50:50 mixture of the VR28-encoding DNA and NNS was introduced at each position. DNA sequence analysis of a random sample of approximately 80 clones revealed an average of six amino acid changes per clone as expected. Lower KDR-Fc concentrations were utilized during selection to favor clones with better affinities to the target. The profile of target binding during the four rounds of selection is shown in FIG. 2. After four rounds of selection the binding population was subcloned and analyzed. Table 5 and FIG. 3A summarize affinity measurements of individual binding clones. The measured binding constants to KDR-Fc ranged from <0.4 to <1.8 nM, a 10-30 improvement over VR28 (11 nM).

[0186] Sequence analysis, some of which is shown in Table 4 (K clones), revealed that while the binding population was diverse, several consensus motifs could be identified among the clones. Most noticeably, Pro87 and Leu84 were found in nearly all clones (as in VR28), suggesting that these residues may be essential for the structure of the binding site. A positively charged amino acid at position 82 appears to be required since only H82K or H82R changes were seen in the sequenced clones and an aliphatic amino acid was predominant at position 78. D81 was often mutated to a G, resulting in the loss of negative charge at this position and a gain in flexibility. In addition, the overall mutation rate in the selected population was comparable to the pool prior to selection, which suggested that the FG loop is very open to changes.

[0187] Several residues in the N-terminus of the .sup.10Fn3 domain of human fibronectin are located in close proximity to the FG loop, as suggested by structural determinations (Main et al, Cell 71:671-678, 1992). The close proximity of the two regions could potentially have a negative impact on target binding. Two incidental mutations in the N-terminal region, L8P and L8Q, resulted in better binding to KDR in a number of selected clones, presumably due to a change of the location of the N-terminus relative to the FG loop. To further test the impact of the N-terminus, we created binding molecules for 23 different KDR binders in which the N-terminal first eight residues before the .beta.-sheet were deleted. We then compared target binding to the non-deleted counterparts. On average, binding to KDR-Fc was about 3-fold better with the deletion, as shown in FIG. 3B.

Example 3

Selection of Binders with Dual Specificities to Human (KDR) and Mouse (Flk-1) VEGFR-2

[0188] VR28 and most of the affinity matured variants (K clones) failed to bind the mouse homolog of KDR, Flk1, as shown in FIG. 4. However, since KDR and Flk1 share a high level of sequence identity (85%, Claffey et al., J. Biol. Chem. 267:16317-16322 (1992), Shima et al., J. Biol. Chem. 271:3877-3883 (1996)), it is conceivable to isolate antibody mimics that can bind both KDR and Flk1. Such dual binders were desirable because they would allow the same molecule to be tested in functional studies in animal models and subsequently in humans.

[0189] The population of clones following FG loop mutagenesis and selection against KDR for four rounds was further selected against Flk1 for an additional three rounds. As shown in FIG. 2 an increase in binding to Flk1 was observed from Round 5 to Round 7, indicating enrichment of Flk1 binders. Analysis of binding for multiple individual clones revealed that in contrast to the clones selected against KDR only (K clones), most clones derived from additional selection against Flk1 (E clones) are able to interact with both KDR and Flk1. The binding constants to both targets, as determined using a radioactive equilibrium binding assay (Table 6 and FIG. 5) and BIAcore (Table 7), indicate that individual clones were able to bind both targets with high affinities.

[0190] For example, E19 has a Kd of 60 pM to KDR, and 340 pM to Flk-1. These results demonstrate that a simple target switch strategy in the selection process, presumably through selection pressures exerted by both targets, has allowed the isolation of molecules with dual binding specificities to both KDR and Flk-1 from a mutagenized population of VR28, a moderate KDR binder that was not able to bind Flk-1. The selected fibronectin-based binding proteins are highly specific to VEGFR-2 (KDR) as no substantial binding to VEGFR1 was observed at high target concentration. Sequence analysis revealed some motifs similar to those observed in the KDR binder pool (Leu and Pro at residues 84 and 87 respectively; positively charged amino acid at residue 82, predominantly Arg) and some that were not maintained (aliphatic at position 78). In addition, the motif ERNGR (residues 78-82) was present in almost all clones binding to Flk-1 (Table 4); this motif was barely discernable in the KDR binding pool. R79 and R82 appear to be particularly important for high affinity binding to Flk-1, since binding to Flk-1, but not KDR, is greatly reduced when a different residue is present at this position (FIG. 6A). To determine the importance of each loop in binding to KDR and Flk-1, the loops of clones E6 and E26 shown in Table 4, were substituted one loop at a time by NNS randomized sequence. As shown in FIG. 6B, after the substitution, the proteins are no longer able to bind either KDR or Flk-1. These results indicate that each loop is required for binding to the targets, suggesting a cooperative participation of all three loops in interacting with the targets.

[0191] An alternative mutagenesis strategy was independently employed to produce clones capable of binding to both targets. The clone 159Q(8)L (Table 4), the product of hypermutagenic PCR affinity maturation of VR28 that binds KDR with high affinity (Kd=2 nM; Table 7) and Flk-1 with poor affinity (Kd>3000 nM), was chosen as a starting point. The first six amino acids of the FG loop were fully randomized (NNS), leaving the following five residues (ELFTP) intact. After six rounds of selection against Flk-1, the binding pool was re-randomized at the DE loop (positions 52-56) and the selection was performed for three additional rounds against Flk-1 and one round against KDR. A number of high affinity binding molecules to both KDR and Flk-1 were thus obtained (Tables 4 and FIG. 4). For example, clone M5FL, while retaining high binding affinity to KDR (Kd=890 pM), can bind Flk-1 at a Kd of 2.1 nM, a 1000-fold improvement over the original clone. Interestingly, the ERNGR motif, found in Flk-1 binding molecules selected from a mutagenized population of VR28, was also present in multiple clones derived from clone 159Q(8)L mutagenesis and selection, despite a full randomization of this region of the FG loop. The isolation of similar binding molecules from two independent libraries suggests that the affinity maturation process is robust for isolating optimal Flk-1 binding motifs located in the FG loop.

Example 4

Cell Surface Binding and Neutralization of VEGF Activity In Vitro

[0192] The functionality of KDR and Flk-1 binding molecules in a cell culture model system was evaluated with E. coli produced binding molecules. Using a detection system consisting of anti-His6 tag murine antibody (the E. coli expressed proteins were expressed with a His tag) and an anti-murine fluorescently labeled antibody the binding molecules were shown to bind specifically to mammalian cells expressing KDR or Flk-1 with low nanomolar EC50s, (FIG. 7 and Table 8).

[0193] More importantly, using recombinant BA/F3 cells (DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH) expressing the extracellular KDR or Flk-1 domain linked to the erythropoietin receptor signaling domain, these molecules inhibited VEGF-stimulated cell proliferation in a dose dependent fashion, with IC50 3-12 nM for KDR expressing cells, and 2-5 nM for Flk-1 expressing cells. The potency of inhibition appears to be similar to control anti-KDR and anti-Flk-1 monoclonal antibodies, as shown in FIG. 8 and Table 9.

[0194] A number of clones were further tested for VEGF-inhibition of the growth of HUVEC cells (Human Umbilical Vein Endothelial Cells). HUVEC cells are natural human cells that are closely related to cells in the body that respond to VEGF. As shown in FIG. 9 and Table 10, the fibronectin-based binding proteins were also active in inhibiting VEGF activity in this human-derived cell system while the wild type fibronectin-based scaffold protein was inactive.

Example 5

Thermal Stability and Reversible Refolding of M5FL Protein

[0195] The thermal stability of KDR-binder M5FL was established using differential scanning calorimetry (DSC). Under standard PBS buffer conditions (sodium phosphate pH 7.4, 150 mM NaCl), M5FL was found to have a single non-reversible thermal melting transition at 56.degree. C. Subsequently, sodium acetate pH 4.5 was identified as a favorable buffer for M5FL protein solubility. DSC experiments in this buffer (100 mM) demonstrated that M5FL is more stable under these conditions (Tm=67-77.degree. C.) and that the melting transition is reversible (FIG. 10). Reversible thermal transitions have been used to identify favorable conditions that support long-term storage of protein therapeutics (Remmele et al, Biochemistry 38:5241 (1999), so Na-acetate pH 4.5 has been identified as an optimized buffer for storing the M5FL protein.

Example 6

In Vitro Binding and Cell-Based Activity of PEGylated M5FL Protein

[0196] The M5FL protein was produced in an E. coli expression system with a C-terminal extension to yield the following protein sequence (C-terminal extension underlined with Cys100 shaded; a significant percentage of protein is produced with the initial methionine removed):

TABLE-US-00010 (SEQ ID NO:199) MGVSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEF TVPLQPPLATISGLKPGVDYTITVYAVTKERNGRELFTPISINYRTEIDK PCQHHHHHH

[0197] The single sulfhydryl of the cysteine residue at position 100 was used to couple to PEG variants using standard maleimide chemistry to yield two different PEGylated forms of M5FL. Both a linear 20 kD PEG and a branched 40 kD PEG (Shearwater Corporation) were conjugated to M5FL to produce M5FL-PEG20 and M5FL-PEG40, respectively. The PEGylated protein forms were purified from unreacted protein and PEG by cation exchange chromatography. Covalent linkage of the two PEG forms of M5FL was verified by SDS-PAGE (FIG. 11) and mass spectroscopy.

[0198] In vitro affinity measurements were made using surface plasmon resonance (SPR) (BIAcore) with both the human and mouse VEGF-receptor target proteins immobilized via amide chemistry on the BIAcore chip. For both target proteins, both the 20 and 40 kD PEGylated M5FL forms were found to have slower on-rates (ka) relative to unmodified M5FL with little effect on off-rates (kd; Table 11).

[0199] The functionality of the PEGylated M5FL preparations was tested using the Ba/F3 system described in Example 4. FIG. 12 shows a plot of A490 (representing the extent of cell proliferation) as a function of concentration of each of the binders. The curves were nearly identical, indicating there was little effect of PEGylation on the biological activity of either of the PEGylated forms.

[0200] The k.sub.on, k.sub.off and K.sub.D were analyzed for a subset of KDR-binding polypeptides and compared to the EC50 for the BaF3 cell-based VEGF inhibition assay. Scatter plots showed that the kon was well-correlated with the EC50, while k.sub.off was poorly correlated. Greater than 90% of KDR-binding proteins with a kon of 105s.sup.-1 or greater had an EC50 of 10 nM or less. K.sub.D is a ratio of kon and koff, and, as expected, exhibits an intermediate degree of correlation with EC50.

[0201] Many of the KDR-binding proteins, including CT-01, were assessed for binding to VEGFR-1, VEGFR-2 and VEGFR-3. The proteins showed a high degree of selectivity for VEGFR-2.

Example 6

Preparation of KDR Binding Protein CT-01 Blocks VEGFR-2 Signaling in Human Endothelial Cells

[0202] Following the methodologies described in the preceding Examples, additional .sup.10Fn3-based KDR binding proteins were generated. As described for the development of the M5FL protein in Example 5, above, proteins were tested for K.sub.D against human KDR and mouse Flk-1 using the BIAcore binding assay and for IC50 in a Ba/F3 assay. A protein termed CT-01 exhibited desirable properties in each of these assays and was used in further analysis.

[0203] The initial clone from which CT-01 was derived had a sequence:

GEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTATISGLK PGVDYTITVYAVTDGWNGRLLSIPISINYRT (SEQ ID NO:200). The FG loop sequence is underlined.

[0204] Affinity maturation as described above produced a core form of CT-01:

TABLE-US-00011 (SEQ ID NO:192) EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTAT ISGLKPGVDYTITVYAVTDGRNGRLLSIPISINYRT.

[0205] The CT-01 molecule above has a deletion of the first 8 amino acids and may include additional amino acids at the N- or C-termini. For example, an additional MG sequence may be placed at the N-terminus. The M will usually be cleaved off, leaving a GEV . . . sequence at the N-terminus. The re-addition of the normal 8 amino acids at the N-terminus also produces a KDR binding protein with desirable properties. The N-terminal methionine is generally cleaved off to yield a sequence:

TABLE-US-00012 (SEQ ID NO:193) VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPPTATISGLKPGVDYTITVYAVTDGRNGRLLSIPISTNYRT.

[0206] For use in vivo, a form suitable for PEGylation may be generated. For example, a C-terminal tail comprising a cysteine was added and expressed, as shown below for a form lacking the eight N-terminal amino acids.

GEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTATISGLK PGVDYTITVYAVTDGRNGRLLSIPISINYRTEIDKPCQ (SEQ ID NO:194). The PEGylated form of this molecule is used in the in vivo experiments described below. A control form with a serine instead of a cysteine was also used:

TABLE-US-00013 (SEQ ID NO:195) GEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTA TISGLKPGVDYTITVYAVTDGRNGRLLSIPISTNYRTEIDKPSQ.

[0207] The same C-terminal tails may also be added to CT-01 forms having the N-terminal eight amino acids, such as is shown in SEQ ID NO:193.

[0208] Additional variants with desirable KDR binding properties were isolated. The following core sequence has a somewhat different FG loop, and may be expressed with, for example, an N-terminal MG sequence, an N-terminal sequence that restores the 8 deleted amino acids, and/or a C-terminal tail to provide a cysteine for PEGylation.

EVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTVPLQPPTATISGLKP GVDYTITVYAVTEGPNERSLFIPISINYRT (SEQ ID NO:196). Another such variant has the core sequence:

TABLE-US-00014 (SEQ ID NO:197) VSDVPRDLEVVAATPTSLLISWRHPHFPTRYYRITYGETGGNSPVQEFTV PLQPPTATISGLKPGVDYTITVYAVTEGPNERSLFIPISINYRT.

[0209] A comparison of these variants shows a consensus sequence for the FG loop of: D/E)GXNXRXXIP (SEQ ID NO:3). With greater particularity, the consensus sequence may be expressed as (D/E)G(R/P)N(G/E)R(S/L)(S/F)IP (SEQ ID NO:4).

Example 7

CT-01 Blocks VEGFR-2 Signaling in Human Endothelial Cells

[0210] As shown in FIG. 13, VEGF-A signaling through VEGFR-2 is mediated by phosphorylation of the intracellular domain of VEGFR-2, followed by activation of pathway involving phospholipase C gamma (PLC.gamma.), Protein Kinase C(PKC), Raf-1, MEK1/2, ERK1/2, leading to endothelial cell proliferation.

[0211] To assess whether KDR binders disclosed herein inhibited activation of this signaling pathway, Human Microvascular Endothelial Cells were treated with a VEGFR binding polypeptide (e.g., CT-01) for 30 min and stimulated with VEGF-A for 5 min. Total cell lysates were analyzed by SDS-PAGE and western analysis, using antibodies specific to phospho-VEGFR-2, non-phospho-VEGFR-2, phosphor-ERK1/2 and non-phospho-ERK1/2.

[0212] As shown in FIG. 13, 130 pM CT-01 inhibits formation of phosphor-VEGFR-2 and also decreases the formation of the downstream phosphorylated ERK1/2. Phosphorylated ERK1/2 is not entirely eliminated, probably due to the fact that ERK1/2 receives signals from a number of additional signaling pathways.

Example 8

Fibronectin-based KDR Binding Proteins Disrupt Signaling by VEGF-A and VEGF-D

[0213] VEGFR-2 is a receptor for three VEGF species, VEGF-A, VEGF-C and VEGF-D.

[0214] Experiments were conducted to evaluate the effects of fibronectin-based KDR binding proteins on VEGF-A and VEGF-D mediated signaling through KDR.

[0215] A Ba/F3 cell line dependent on Flk-1 mediated signaling was generated. As shown in the left panel of FIG. 14, cell viability could be maintained by treating the cells with VEGF-A or VEGF-D, although significantly higher levels of VEGF-D were required.

[0216] As shown in the middle panel of FIG. 14, cells were maintained in the presence of 15 ng/ml of VEGF-A and contacted with the M5FL or CT-01 proteins disclosed herein, or with the DC-101 anti-Flk-1 antibody. Each reagent reversed the VEGF-A-mediated cell viability, indicating that VEGF-A signaling through Flk-1 was blocked.

[0217] As shown in the right panel of FIG. 14, cells were maintained in the presence of 300 ng/ml of VEGF-D and contacted with the M5FL or F10 proteins disclosed herein, or with an anti-VEGF-A antibody. M5FL and F10 reversed the VEGF-D-mediated cell viability, indicating that VEGF-D signaling through Flk-1 was blocked. The anti-VEGF-A antibody had no effect, demonstrating the specificity of the assay.

Example 9

Pharmacokinetics

[0218] Pharmacokinetic Studies: Native CT-01 or a pegylated form (40 kDa PEG, CT-01PEG40) were iodinated with .sup.125I. 10-20 mCi of iodinated proteins were injected into adult male rats either i.v. or i.p. and iodinated proteins levels were determined at the indicated times. For tissue distribution studies, rats were sacrificed at 15 min, 2 hr and 6 hr and radioactivity levels determined. See FIGS. 15 and 16. Unmodified CT-01 is a 12 kDa protein that is rapidly cleared from the blood. The area-under-curve value (AUC) value is 14.6 hr*mg/mL with a clearance of 69.9 mL/hr/kg, a maximum serum concentration of 9.1 mg/ml. The initial half-life (a) is 0.3 hours and the second phase half-life (.beta.) is 13.5 hours. By comparison, i.v. PEGylated CT-01 has greatly increased presence in the blood, mostly because of a dramatic decrease in the initial phase of clearance. The AUC is increased greater than 10 fold to 193, the clearance rate is decreased by greater than 10 fold to 5.2, the Cmax is 12.9 mg/mL. The .alpha. half-life is increased to 1 hour, and the .beta. is increased to 16.2 hours. These pharmacokinetics in rats are equivalent to a twice-weekly dosing regimen in humans, a rate of dosing that is well within acceptable ranges.

[0219] Intraperitoneal (i.p.) administration of PEGylated CT-01 had reservoir-like pharmacokinetics. There was no initial spike in the blood concentration of CT-01. Instead, the amount of CT-01 built up more slowly and decreased slowly. Such pharmacokinetics may be desirable where there is concern about side effects from the initial spike in CT-01 concentration upon intravenous administration. It is likely that other .sup.10FN3-based agents would exhibit similar behavior in i.p. administration.

[0220] Accordingly, this may be a generalizable mode for achieving a time-delayed dosing effect with .sup.10FN3-based agents.

[0221] As shown in FIG. 16, the liver is the primary route for secretion of the PEGylated form of CT-01. No long term accumulation of CT-01 was detected.

[0222] Similar results were obtained using a CT-01 conjugated to a 20 kDa PEG moiety.

Example 10

In Vivo Efficacy of CT-01

[0223] The Miles assay, as outlined in FIG. 17, is used to evaluate Dose, Schedule and Administration parameters for the tumor efficacy studies. Balb/c female mice were injected i.p. with buffer or CT-01PEG40 at 1, 5 and 20 mg/kg 4 hr prior to VEGF challenge. Intradermal focal administration of VEGF-A into the back skin induces vessel leakage of Evans blue dye (FIGS. 17 and 18).

[0224] Mice treated with a KDR binding agent showed a statistically significant decrease in the level of VEGF-mediated vessel leakage. Both 5 mg/kg and 20 mg/kg dosages with CT-01 showed significant results. Therefore, a 5 mg/kg dosage was selected for mouse tumor model studies.

Example 11

CT-01 Inhibits Tumor Growth

B 16-F10 Murine Melanoma Tumor Assay:

[0225] 2.times.10.sup.6 B16-F10 murine melanoma tumor cells were implanted subcutaneously into C57/BL male mice at Day 1. At day 6 a palpable mass was detected. On day 8 when tumors were of measurable size, daily i.p. injections of either Vehicle control, 5, 15, or 40 mg/kg CT-01PEG40 were started. The lowest dose 5 mg/kg decreased tumor growth.

[0226] At day 18, mice treated with 15 and 40 mg/kg showed 50% and 66% reduction in tumor growth. See FIG. 19.

U87 Human Glioblastoma Assay:

[0227] 5.times.10.sup.6 U87 human glioblastoma tumor cells were implanted subcutaneously into nude male mice. When tumor volume reached approximately 50 mm3 treatment started (day 0). Vehicle control, 3, 10, or 30 mg/kg CT-01PEG40 were injected i.v. every other day (EOD). The anti-Flk-1 antibody DC101 was injected at 40 mg/kg twice a week as published for its optimal dose schedule. The lowest dose 3 mg/kg decreased tumor growth. At day 12, mice treated with 10 and 30 mg/kg showed 50% reduction in tumor growth. See FIG. 20. Effectiveness is comparable to that of the anti-Flk-1 antibody.

[0228] The following materials and methods were used for the experiments described in Examples 1-11.

Recombinant Proteins:

[0229] Recombinant human VEGF.sub.165, murine VEGF.sub.164, human neurotrophin-4 (NT4), human and mouse vascular endothelial growth factor receptor-2 Fc chimeras (KDR-Fc and Flk-1-Fc) were purchased from R&D systems (Minneapolis, Minn.). Biotinylation of the target proteins was carried out in 1.times.PBS at 4.degree. C. for 2 hours in the presence of EZ-Lik.TM. Sulfo-NHS-LC-LC-Biotin (Pierce, Ill.). Excess of EZ-Link.TM. Sulfo-NHS-LC-LC-Biotin was removed by dialysis against 1.times.PBS. The level of biotinylation was determined by mass spectroscopy and target protein concentrations were determined using Coomassie Protein Plus Assay (Pierce, Ill.).

Primers:

[0230] The following oligonucleotides were prepared by chemical synthesis for eventual use in library construction and mutagenesis of selected clones.

TABLE-US-00015 T7 TMV Fn: 5' GCG TAA TAC GAC TCA CTA TAG GGA CAA TTA CTA TTT ACA ATT ACA ATG GTT TCT GAT GTT CCG AGG 3' (SEQ ID NO:201) T7 TMV N-terminus deletion: 5' GCG TAA TAC GAC TCA CTA TAG GGA CAA TTA CTA TTT ACA ATT ACA ATG GAA GTT GTT GCT GCG ACC CCC ACC AGC CTA 3' (SEQ ID NO:202) MK165-4 A20: 5' TTT TTT TTT TTT TTT TTT TTA AAT AGC GGA TGC CTT GTC GTC GTC GTC CTT GTA GTC 3' (SEQ ID NO:203) N-terminus forward: 5' ATG GTT TCT GAT GTT CCG AGG GAC CTG GAA GTT GTT GCT GCG ACC CCC ACC AGC CTA CTG ATC AGC TGG 3' (SEQ ID NO:204) BCDE reverse: 5' AGG CAC AGT GAA CTC CTG GAC AGG GCT ATT TCC TCC TGT TTC TCC GTA AGT GAT CCT GTA ATA TCT 3' (SEQ ID NO:205) BCDE forward: 5' AGA TAT TAC AGG ATC ACT TAC GGA GAA ACA GGA GGA AAT AGC CCT GTC CAG GAG TTC ACT GTG CCT 3' (SEQ ID NO:206) DEFG reverse: 5' AGT GAC AGC ATA CAC AGT GAT GGT ATA ATC AAC TCC AGG TTT AAG GCC GCT GAT GGT AGC TGT 3' (SEQ ID NO:207) DEFG forward: 5' ACA GCT ACC ATC AGC GGC CTT AAA CCT GGA GTT GAT TAT ACC ATC ACT GTG TAT GCT GTC ACT 3' (SEQ ID NO:208) C-terminus polyA: 5' TTT TTT TTT TTT TTT TTT TAA ATA GCG GAT GCC TTG TCG TCG TCG TCC TTG TAG TCT GTT CGG TAA TTA ATG GAA AT 3' (SEQ ID NO:209) Hu3'FLAGSTOP: 5' TTT TAA ATA GCG GAT GCC TTG TCG TCG TCG TCC TTG TAG TCT GTT CGG TAA TTA ATG G 3' (SEQ ID NO:210) R28FG-50: 5' GTG TAT GCT GTC ACT 123 145 463 665 165 465 163 425 625 645 447 ATT TCC ATT AAT TAC 3' , (SEQ ID NO:211), where 1 = 62.5%G + 12.5%A + 12.5% T + 12.5%C; 2 = 2.5%G + 12.5%A + 62.5%T + 12.5%C; 3 = 75%G + 25%C; 4 = 12.5%G + 12.5%A + 12.5%T + 62.5%C; 5 = 25%G + 75%C; 6 = 12.5%G + 62.5%A + 12.5%T + 12.5%C; 7: 25%G + 50%A + 25%C F1U2: 5' TAA TAC GAC TCA CTA TAG GGA CAA TTA CTA TTT ACA ATT CTA TCA ATA CAA TGG TGT CTG ATG TG CCG 3' (SEQ ID NO:212) F2: 5' CCA GGA GAT CAG CAG GGA GGT CGG GGT GGC AGC CAC CAC TTC CAG GTC GCG CGG CAC ATC AGA CAC CAT TGT 3' (SEQ ID NO:213) F3159: 5' ACC TCC CTG CTG ATC TCC TGG CGC CAT CCG CAT TTT CCG ACC CGC TAT TAC CGC ATC ACT TAC G 3' (SEQ ID NO:214) F4: 5' CAC AGT GAA CTC CTG GAC CGG GCT ATT GCC TCC TGT TTC GCC GTA AGT GAT GCG GTA ATA GCG 3' (SEQ ID NO:215) F5159: 5' CGG TCC AGG AGT TCA CTC TGC CGC TGC AGC CGC CGG CGG CTA CCA TCA GCG GCC TTA AAC C 3' (SEQ ID NO:216) F5-X5: 5' CG GTC CAG GAG TTC ACT GTG CCG NNS NNS NNS NNS NNS GCT ACC ATC AGC GGC CTT AAA CC 3' (SEQ ID NO:217) F6: 5' AGT GAC AGC ATA CAC AGT GAT GGT ATA ATC AAC GCC AGG TTT AAG GCC GCT GAT GGT AG 3' (SEQ ID NO:218) F7X6159: 5' ACC ATC ACT GTG TAT GCT GTC ACT NNS NNS NNS NNS NNS NNS GAA CTG TTT ACC CCA ATT TCC ATC AAC TAC CGC ACA GAC TAC AAG 3' (SEQ ID NO:219) F8: 5' AAA TAG CGG ATG CGC GTT TGT TCT GAT CTT CCT TAT TTA TGT GAT GAT GGT GGT GAT GCT TGT CGT CGT CGT CCT TGT AGT CTG TGC GGT AGT TGA T 3' (SEQ ID NO:220) G2asaiA20: 5' TTT TTT TTT TTT TTT TTT TTA AAT AGC GGA TGC GCG TTT GTT CTG ATC TTC 3' (SEQ ID NO:221) C2RT: 5' GCG CGT TTG TTC TGA TCT TCC 3' (SEQ ID NO:222) hf01 BC reverse: 5' TGCC TCC TGT TTC GCC GTA AGT GAT GCG GTA ATA GCG SNN SNN SNN SNN SNN SNN SNN CCA GCT GAT CAG CAG 3' (SEQ ID NO:223) hf01 DE reverse: 5' GAT GGT AGC TGT SNN SNN SNN SNN AGG CAC AGT GAA CTC CTG GAC AGG GCT ATT GCC TCC TGT TTC GCC 3' (SEQ ID NO:224) hf01 FG reverse: 5' GT GCG GTA ATT AAT GGA AAT TGG SNN SNN SNN SNN SNN SNN SNN SNN SNN SNN AGT GAC AGC ATA CAC 3' (SEQ ID NO:225) BCDE rev: 5' CCT CCT GTT TCT CCG TAA GTG 3' (SEQ ID NO:226) BCDEfor: 5' CAC TTA CGG AGA AAC AGG AGG 3' (SEQ ID NO:227) hf01 DE-FG forward: 5' ACA GCT ACC ATC AGC GGC CTT AAA CCT GGC GTT GAT TAT ACC ATC ACT GTG TAT GCT GTC ACT 3' (SEQ ID NO:228) Front FG reverse: 5' AGT GAC AGC ATA CAC AGT 3' (SEQ ID NO:229) hf01 RT Flag PolyA reverse: 5' TTT TTT TTT TTT TTT TTT TTA AAT AGC GGA TGC CTT GTC GTC GTC GTC CTT GTA GTC TGT GCG GTA ATT AAT GGA 3' (SEQ ID NO:230) 5-RI-hKDR-1B: 5' TAG AGA ATT CAT GGA GAG CAA GGT GCTG 3' (SEQ ID NO:231) 3-EPO/hKDR-2312B: 5' AGG GAG AGC GTC AGG ATG AGT TCC AAG TTC GTC TTT TCC 3' (SEQ ID NO:232) 5-RI-mKDR-1: 5' TAG AGA ATT CAT GGA GAG CAA GGC GCT G 3' (SEQ ID NO:233) 3-EPO/mKDR-2312: 5' AGG GAG AGC GTC AGG ATG AGT TCC AAG TTG GTC TTT TCC 3' (SEQ ID NO:234) 5-RI-hTrkB-1: 5' TAG AGA ATT CAT GAT GTC GTC CTG GAT AAG GT 3' (SEQ ID NO:235) 3-EpoR/hTrkB-1310: 5' AGG GAG AGC GTC AGG ATG AGA TGT TCC CGA CCG GTT TTA 3' (SEQ ID NO:236) 5-hKDR/EPO-2274B: 5' GGA AAA GAC GAA CTT GGA ACT CAT CCT GAC GCT CTC CCT 3' (SEQ ID NO:237) 5-mKDR/EPO-2274: 5' GGA AAA GAC CAA CTT GGA ACT CAT CCT GAC GCT CTC CCT 3' (SEQ ID NO:238) 3-XHO-EpoR-3066: 5' TAG ACT CGA GTC AAG AGC AAG CCA CAT AGCT 3' (SEQ ID NO:239) 5'hTrkB/EpoR-1274: 5' TAA AAC CGG TCG GGA ACA TCT CAT CCT GAC GCT CTC CCT 3' (SEQ ID NO:240)

Buffers

[0231] The following buffers were utilized in the experiments described herein. Buffer A (100 mM Tris HCl, 1M NaCl, 0.05% Tween-20, pH 8.0); Buffer B (1.times.PBS, 0.02% Triton X100); Buffer C (100 mM Tris HCl, 60 mM EDTA, 1M NaCl, 0.05% Triton X100, pH 8.0); Buffer Ca (100 mM Tris HCl, 1M NaCl, 0.05% Triton X100, pH 8.0); Buffer D (2M NaCl, 0.05% Triton); Buffer E (1.times.PBS, 0.05% Triton X100, pH 7.4); Buffer F (1.times.PBS, 0.05% Triton X100, 100 mM imidazole, pH 7.4); Buffer G (50 mM HEPES, 150 mM NaCl, 0.02% TritonX-100, 1 mg/ml bovine serum albumin, 0.1 mg/ml salmon sperm DNA, pH 7.4); Buffer H (50 mM HEPES, 150 mM NaCl, 0.02% TritonX-100, pH 7.4); Buffer I (1.times.PBS, 0.02% TritonX-100, 1 mg/ml bovine serum albumin, 0.1 mg/ml salmon sperm DNA, pH 7.4); Buffer J (1.times.PBS, 0.02% TritonX-100, pH 7.4); Buffer K (50 mM NaH.sub.2PO.sub.4, 0.5 M NaCl, 5% glycerol, 5 mM CHAPS, 25 mM imidazole, 1.times. Complete.TM. Protease Inhibitor Cocktail (Roche), pH 8.0); Buffer L (50 mM NaH.sub.2PO.sub.4, 0.5 M NaCl, 5% glycerol, 25 mM imidazole, pH 8.0); Buffer M (1.times.PBS, pH 7.4, 25 mM imidazole, pH 7.4); Buffer N (1.times.PBS, 250 mM imidazole, pH 7.4); Buffer 0 (10 mM HEPES, 150 mM NaCl, 0.005% Tween 20, pH 7.4).

Primary Library Construction:

[0232] The construction of the library using the tenth domain of human fibronectin as a scaffold was previously described (Xu et al, 2002, supra). Three loop regions, corresponding to positions 23-29, 52-55, and 77-86, respectively, were randomized using NNS (standard nucleotide mixtures, where N=equimolar mixture of A, G, T, C; S=equimolar mixture of G and C) as the coding scheme. Similar libraries were constructed containing randomized regions only at positions 23-29 and 77-86 (two loop library) or only at positions 77-86 (one loop library). These libraries were mixed in approximately equimolar amounts. This mixed library contained .about.1.times.10.sup.13 clones and was used in the KDR selection that identified VR28.

Mutagenic Library Construction:

[0233] Hypermutagenic PCR. Scaffold mutation T(69)I in VR28 clone was corrected back to wild type sequence by PCR (see below) and no change in binding characteristics of VR28 binder to KDR was observed. Mutations were introduced into the loop regions of VR28 using conditions described previously (Vartanian et al, Nuc. Acid Res. 24:2627-2631, 1996). Three rounds of hypermutagenic PCR were conducted on a VR28 template using primer pairs flanking each loop (N-terminus forward/BCDE reverse, BCDE forward/DEFG reverse, DEFG forward/C-terminus polyA). The resulting fragments were assembled using overlap extension and PCR with flanking primers T7TMV Fn and MK165-4 A20. DNA sequencing of the clones from the final PCR reaction confirmed correct assembly of the library. Up to 30% mutagenesis rate was observed in the loop regions, as compared to 1.5% in the scaffold regions.

[0234] Oligo mutagenesis. Oligo mutagenesis of the FG loop of VR28 by PCR utilized the VR28FG-50 primer, DEFG reverse primer and flanking primers. At each nucleotide position encoding the FG loop, primer VR28FG-50 contained 50% of the VR28 nucleotide and 50% of an equimolar mixture of all four nucleotides (N) or of G or C(S). This scheme was designed to result in approximately 67% of the amino acids of the VR28 FG loop being randomly replaced by another amino acid which was confirmed by DNA sequencing.

[0235] 159 (Q8L) randomized sub-libraries. Oligo mutagenesis of the FG loop of Clone 159 (Q8L) clone, a three-step extension and amplification was performed. For the first extension, pairs of primers (a: F1U2/F2, b: F3159/F4, c: F5159/F6, d: F7X6159/F8) were mixed in equal concentrations (100 pmol each) and amplified for 10 cycles. For the second extension, 1/20 of the first reactions were combined (a/b and c/d) and amplification was continued for another 10 cycles. To bias the amplification in favor of extension rather than re-annealing of fully complementary fragments, a linear amplification of the half-construct products (0.5 pmol each) was performed for an additional 20 cycles using 50 pmol of either F1U2 forward primer for fragment ab, or the C2asaiA20 reverse primer for fragment cd. Finally, the extended half-construct fragments ab and cd were combined and amplified for 20 cycles without any additional components. Primer F7X6159 contained NNS at each of the first 6 coding positions of Clone 159 (Q8L) and was otherwise identical to Clone 159 (Q8L). Correct assembly of the library 159 (Q8L)-FGX6 was confirmed by DNA sequencing of clones from the final PCR reaction. The sub-library contained .about.1.times.10.sup.9 clones.

[0236] For randomization of the DE loop of post round 6 (PR6) selection pool of the 159 (Q8L)-FGX6 library, two half-construct fragments were prepared by PCR using primers F1U2/F4 and F5X5/C2asaiA20. The F5X5 primer contained NNS at the four positions of the DE loop as well as at position 56. Then, the extended fragments ab and cd were combined and amplified for 20 cycles without any additional components.

[0237] Introduction of point mutations, deletion and random (NNS) loop sequences into fibronectin-based scaffold proteins:

Scaffold mutation T(69)I of VR28 binder was corrected back to wild type sequence in two-step PCR using VR28 clone as a template. Half-construct fragments, obtained with primers N-terminus forward/DEFG reverse and DEFG forward/C-terminus polyA, were combined and the whole VR28 (169T) clone (designated as VR28 in the text) was constructed using primers T7TMV Fn and MK165-4 A20. Correction of N-terminus mutations in clone 159 (Q8 to L) was performed by PCR with primers N-terminus forward/C-terminus polyA followed by extension with primers T7TMV Fn and MK165-4 A20.

[0238] Introduction of deletion .DELTA.1-8 into the N-terminus of fibronectin-based scaffold proteins was performed by amplification using primers T7 TMV N-terminus deletion and MK165-4 A20.

[0239] Construction of the chimeras of E clones containing NNS loop sequences was performed by two-step PCR. Loop regions were amplified using primers T7 TMV N-terminus deletion/BCDE rev (a: BC loop of E clones); N-terminus forward/hf01 BC reverse (b: BC NNS); BCDE for/Front FG reverse (c: DE loop of E clones); BCDE for/hf01 DE reverse (d: DE NNS); hf01 DE-FG forward/hf01 RT-Flag PolyA reverse (e: FG of E clones); hf01 DE-FG forward/hf01 FG reverse (f: FG NNS). Fragments b/c/e, a/d/e, a/c/f were combined and the whole pools were constructed by extension and amplification using primers T7Tmv N-terminus deletion and hf01 RTFlag PolyA reverse.

[0240] All constructs were verified and/or analyzed by DNA sequencing. All constructs and mutagenic libraries contained T7 TMV promoter at the 5' flanking region and Flag tag or His.sub.6 tag sequences at 3' flanking region for RNA-protein fusion production and purification in vitro.

RNA-Protein Fusion Production

[0241] For each round of selection PCR DNA was transcribed using MegaScript transcription kit (Ambion) at 37.degree. C. for 4 hours. Template DNA was removed by DNase I (Ambion) digestion at 37.degree. C. for 20 minutes. RNA was purified by phenol/chloroform extraction followed by gel filtration on a NAP-25 column (Amersham). The puromycin linker PEG 6/10 (5' Pso u agc gga ugc XXX XXX CC Pu 3', where Pso=C6-Psoralen, u,a,g,c=2'OMe-RNA, C=standard amidities, X: Spacer Phosphoramidite 9 (9-O-Dimethoxytrityl-triethylene glycol, 1-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite); Pu=Puromycin-CPG) was synthesized as described previously (Kurz et al, Nuc. Acid Res. 28:83, 2000). The linker was annealed to the library RNA in 0.1 M NaCl, 25 mM Tris HCl, pH 7.0, by gradient temperature decrease from 85.degree. C. to 4.degree. C. The linker and RNA were then cross linked by exposing to UV light (365 nm) for 15 minutes. The cross-linked mixture (600 pmol RNA) was included in an in vitro translation reaction using rabbit reticulocyte lysate translation kit (Ambion) in the presence of .sup.35S-labeled methionine at 30.degree. C. for 60 minutes. To enhance fusion formation, 0.5 M KCl and 0.05 M MgCl.sub.2 were added to the reaction and incubated for 30 minutes at 4.degree. C. Fusion molecules were purified using oligo-dT cellulose (Sigma) chromatography as follows. The translation and fusion mix was diluted into buffer A (100 mM Tris HCl, 1M NaCl, 0.05% Tween-20, pH 8.0) and added to oligo dT cellulose. The slurry was rotated at 4.degree. C. for 1 hour and transferred to a spin column. Oligo dT cellulose beads were washed on the column with 10 column volumes of buffer A and eluted with 3 column volumes of H.sub.2O. Reverse transcription reaction was conducted with SuperScript II Reverse Transcription kit (Invitrogen) for 1 hour at 42.degree. C. using primer Hu3'FLAGSTOP. To decrease potential non-specific binding through reactive cysteines the thiol groups were reacted with 1 mM of 2-nitro-5-thiocyanatobenzoic acid (NTCB) or N-ethylmaleimide (NEM) alternatively over the course of the selection. The reaction was carried out for 1 hour at room temperature. Fusion molecules were further purified by anti-FLAG affinity chromatography using M2 agarose (Sigma). The M2 beads were added to the reaction and rotated in buffer B (1.times.PBS, 0.02% Triton X100) for 1 hour at 4.degree. C. Then the beads were applied to a spin column, washed with 5 column volumes of buffer B and fusion molecules were eluted with 3 column volumes of 100 .mu.M Flag peptide DYKDDDDK (Sigma) in buffer G. Fusion yield was calculated based on specific activity measured by scintillation counting of .sup.35S-methionine in the samples.

[0242] For the 159 (Q8L) randomized library, RNA-protein fusion was prepared using a streamlined, semi-automated procedure in a Kingfisher.TM. (ThermoLabSystems). The steps were similar to the procedure described above except for several steps described below. Purification of the RNA-protein fusion molecules was performed in buffer C (100 mM Tris HCl, 60 mM EDTA, 1M NaCl, 0.05% Triton X100, pH 8.0) on magnetic oligo dT beads (GenoVision). The beads were washed with 10 reaction volumes of buffer Ca (100 mM Tris HCl, 1M NaCl, 0.05% Triton X100, pH 8.0) and fusion proteins were eluted with one volume of H.sub.2O. Reverse transcription (RT) was conducted using primer C2RT. Fusion proteins were further purified by His-tag affinity chromatography using Ni-NTA magnetic beads (Qiagen). The RT reaction was incubated with Ni-NTA beads in buffer D (2M NaCl, 0.05% Triton) for 20 minutes at room temperature, the beads were then washed with 10 reaction volumes of buffer E (1.times.PBS, 0.05% Triton X100, pH 7.4) and fusion molecules were eluted with one volume of buffer F (1.times.PBS, 0.05% Triton X100, 100 mM imidazole, pH 7.4).

Selection:

[0243] Primary selection against KDR. Fusion library (.about.10.sup.13 clones in 1 ml) was incubated with 150 .mu.l of Protein A beads (Dynal) which was pre-immobilized with 200 nM of human IgG1 for 1 hour at 30.degree. C. prior to selection to reduce non-specific binding to both Protein A baeds and Fc protein (preclear). The supernatant was then incubated in buffer G (50 mM HEPES, 150 mM NaCl, 0.02% TritonX-100, 1 mg/ml bovine serum albumin, 0.1 mg/ml salmon sperm DNA, pH 7.4) with KDR-Fc chimera for 1 hour at 30.degree. C. with end-over-end rotation. Final concentrations of KDR-Fc were 250 nM for Round 1, 100 nM for rounds 2-4 and 10 nM for rounds 5 and 6. The target was captured on 300 .mu.l of Protein A beads (Round 1) or 100 .mu.l of Protein A beads (Rounds 2-6) for 30 minutes at 30.degree. C. with end-over-end rotation and beads were washed 5 times with 1 ml of buffer G (50 mM HEPES, 150 mM NaCl, 0.02% TritonX-100, pH 7.4). Bound fusion molecules were eluted with 100 .mu.l of 0.1 M KOH into 50 .mu.l of 1 M Tris HCl, pH 8.0. DNA was amplified from elution by PCR using flanking primers T7TMV Fn and MK165-4 A20.

[0244] Affinity and specificity maturation of KDR binder VR28. Clone VR28 was mutagenized by hypermutagenic PCR or oligo-directed mutagenesis as described above and fusion sub-libraries were constructed. Following pre-clear with Protein A beads selection was performed in buffer I (1.times.PBS, 0.02% TritonX-100, 1 mg/ml bovine serum albumin, 0.1 mg/ml salmon sperm DNA, pH 7.4) for four rounds according to procedure described above. DNA was amplified from elution by PCR using primers T7TMV Fn and MK165-4 A20. Lower target concentrations (0.1 nM KDR for first four rounds of selection) were used for libraries derived from oligo mutagenesis and then 1 nM mouse VEGF-R2 (Flk-1) was introduced for three additional rounds of selection. Primers T7 TMV N-terminus deletion and MK165-4 A20 were used for PCR in the last 3 rounds. For specificity maturation of KDR binder 159 first 6 positions of the FG loop of clone 159 Q(8)L were randomized by PCR as described above. Binding of the fusion sub-library to biotinylated mouse VEGF-R2 (70 nM) was performed in buffer I at room temperature for 30 minutes. The rest of the selection procedure was continued in Kingfisher.TM. (ThermoLabSystems). The biotinylated target was captured on 50 .mu.l of streptavidin-coated magnetic beads (Dynal) and the beads were washed with 10 volumes of buffer I and one volume of buffer J (1.times.PBS, 0.02% TritonX-100, pH 7.4). Bound fusion molecules were eluted with 100 .mu.l of 0.1 M KOH into 5.01 of 1 M Tris HCl, pH 8.0. DNA was amplified from elution by PCR using primers F1U2 and C2asaiA20. After four rounds of selection an off-rate/rebinding selection against 7 nM Flk-1 was applied for another two rounds as follows. After the binding reaction with biotinylated mouse Flk-1 had progressed for 30 minutes, a 100-fold excess of non-biotinylated Flk-1 was added and the reaction continued for another 6 hours to allow time for the weak binders to dissociate. The biotinylated target was captured on 50 .mu.l of streptavidin beads (Dynal) and beads were washed 5 times with 1 ml of buffer J. Bound fusion molecules were eluted by incubation at 75.degree. C. for 5 minutes. Supernatant was subjected to re-binding to 7 nM Flk-1 and standard selection procedure was continued. DNA from the final elution pool was subjected to DE loop randomization (see above) and fusion sub-library was selected against 7 nM mouse VEGF-R2 for three rounds. At the fourth round an off-rate selection was applied with re-binding to 1 nM human VEGF-R2. Final DNA was amplified from elution by PCR using primers F1U2 and C2asaiA20.

Radioactive Equilibrium Binding Assay

[0245] To prepare .sup.35S-labeled binding proteins for analysis, mRNA was prepared as described above for RNA-protein fusion production but the linker ligation step was omitted. The mRNA was expressed using rabbit reticulocyte lysate translation kit (Ambion) in the presence of .sup.35S-labeled Met at 30.degree. C. for 1 hour. Expressed protein was purified on M2-agarose Flag beads (Sigma) as described above. This procedure produced the encoded protein without the nucleic acid tail. In a direct binding assay, VEGF-R2-Fc fusions in concentrations ranging from 0 to 200 nM were added to a constant concentration of the purified protein (0.2 or 0.5 nM) and incubated at 30.degree. C. for 1 hour in buffer B. The receptor-binder complexes were captured using Protein A magnetic beads for another 10 minutes at room temperature using a Kingfisher.TM.. The beads were washed with six reaction volumes of buffer B. The protein was eluted from the beads with 100 .mu.L of 0.1 M KOH. 50 .mu.L of the reaction mixture and elution were dried onto a LumaPlate-96 (Packard) and the amount of .sup.35S on the plate was measured using a TopCount NXT instrument (Packard). The amount of fibronectin-based scaffold protein bound to the target was estimated as a percent of radioactivity eluted from Protein A magnetic beads compare to radioactivity in the reaction mixture. Nonspecific binding of fibronectin-based scaffold proteins to the beads was determined by measuring binding in the absence of KDR-Fc and represented less than 1-2% of the input. Specific binding was obtained through subtraction of nonspecific binding from total binding. Data was analyzed using the GraphPad Prizm software (GraphPad Software, Inc, San Diego, Calif.), fitted using a one site, non-linear binding equation.

Expression and Purification of Soluble Fibronectin-Based Scaffold Protein Binders:

[0246] For expression in E. coli residues 1-101 of each clone followed by the His.sub.6 tag were cloned into a pET9d-derived vector and expressed in E. coli BL21 (DE3) pLysS cells (Invitrogen). 20 ml of overnight culture was used to inoculate 1 liter of LB medium containing 50 .mu.g/mL kanamycin and 34 .mu.g/mL chloromphenicol. The culture was grown at 37.degree. C. until A.sub.600 0.4-0.6. After induction with 1 mM isopropyl-.beta.-thiogalactoside (IPTG, Invitrogen) the culture was grown for another 3 hours at 37.degree. C. and harvested by centrifugation for 30 minutes at 3,000 g at 4.degree. C. The cell pellet was resuspended in 50 mL of lysis buffer K (50 mM NaH.sub.2PO.sub.4, 0.5 M NaCl, 5% glycerol, 5 mM CHAPS, 25 mM imidazole, 1.times. Complete.TM. Protease Inhibitor Cocktail (Roche), pH 8.0) Buffer L and sonicated on ice at 80 W for four 15 second pulses separated by ten-second pauses. The soluble fraction was separated by centrifugation for 30 minutes at 30,000 g at 4.degree. C. The supernatant was rotated for 1 hour at 4.degree. C. with 10 mL of TALON.TM. Superflow.TM. Metal Affinity Resin (Clontech) pre-equilibrated with wash buffer L (50 mM NaH.sub.2PO.sub.4, 0.5 M NaCl, 5% glycerol, 25 mM imidazole, pH 8.0). The resin was then washed with 10 column volumes of buffer L and 30 column volumes of buffer M (1.times.PBS, pH 7.4, 25 mM imidazole, pH 7.4). Protein was eluted with 5 column volumes of buffer N (1.times.PBS, 250 mM imidazole, pH 7.4) and dialyzed against 1.times.PBS at 4.degree. C. Any precipitate was removed by filtering at 0.22 .mu.m (Millipore).

BIAcore Analysis of the Soluble Fibronectin-Based Scaffold Proteins:

[0247] The binding kinetics of fibronectin-based scaffold proteins binding proteins to the target was measured using BIAcore 2000 biosensor (Pharmacia Biosensor). Human and mouse VEGF-R2-Fc fusions were immobilized onto a CM5 sensor chip and soluble binding proteins were injected at concentrations ranging from 0 to 100 nM in buffer 0 (10 mM HEPES, 150 mM NaCl, 0.005% Tween 20, pH 7.4). Sensorgrams were obtained at each concentration and were evaluated using a program, BIA Evaluation 2.0 (BIAcore), to determine the rate constants k.sub.a (k.sub.on) and k.sub.d (k.sub.off) The affinity constant, K.sub.D was calculated from the ratio of rate constants k.sub.off/k.sub.on.

[0248] For inhibition experiments, human VEGF.sub.165 was immobilized on a surface of CM-5 chip and KDR-Fc was injected at a concentration of 20 nM in the presence of different concentrations of soluble binding proteins ranging from 0 to 100 nM. IC.sub.50 was determined at a concentration when only 50% of KDR-Fc binding to the chip was observed.

Reversible Refolding of a VEGFR Binding Polypeptide:

[0249] Differential scanning calorimetry (DSC) analysis was performed on M5FL protein in 100 mM sodium acetate buffer (pH 4.5). An initial DSC run (Scan 1) was performed in a N-DSC II calorimeter (Calorimetry Sciences Corp) by ramping the temperature from 5-95.degree. C. at a rate of 1 degree per minute, followed by a reverse scan (not shown) back to 10 degrees, followed by a second run (Scan 2). Under these conditions, data were best fit using a two transition model (Tm=77.degree. C. and 67.degree. C. using Orgin software (OrginLab Corp)). See FIG. 10.

PEGylation of the M5FL Protein:

[0250] The C100-form of the M5FL protein, which has the complete sequence of M5FL with the Ser at position 100 mutated to a Cysteine including the additional C-terminal His-tag used to purify the protein. The purified M5FL-C100 protein was modified at the single cysteine residue by conjugating various maleimide-derivatized PEG forms (Shearwater). The resulting reacted proteins were run on a 4-12% polyacrylamide gel (FIG. 11).

Construction of Cell Lines:

[0251] Plasmid construction. Plasmids, encoding chimeric receptors composed of the transmembrane and cytoplasmic domains of the human erythropoietin receptor (EpoR) fused to the extracellular domains of KDR, Flk-1, or human TrkB were constructed by a two-step PCR procedure. PCR products encoding the extracellular domains were amplified from plasmids encoding the entire receptor gene: KDR (amino acids 1 to 764) was derived from clone PR1371_H11 (OriGene Technologies, Rockville, Md.) with primers 5-RI-hKDR-1B/3-EPO/hKDR-2312B, flk-1 (amino acids 1 to 762) was derived from clone #4238984 (IMAGE) with primers 5-RI-mKDR-1/3-EPO/mKDR-2312, and human TrkB (from amino acids 1 to 430) from clone #X75958 (Invitrogen Genestorm) with primers 5-RI-hTrkB-1/3-EpoR/hTrkB-1310. PCR products encoding the EpoR transmembrane and cytoplasmic domains (amino acids 251 to 508) were amplified from clone #M60459 (Invitrogen Genestorm) with the common primer 3-XHO-EpoR-3066 and one of three gene-specific primers 5-hKDR/EPO-2274B (KDR), 5-mKDR/EPO-2274 (flk-1), and 5'hTrkB/EpoR-1274 (human TrkB), which added a short sequence complementary to the end of the receptor fragment PCR product. Second, PCR products encoding the two halves of the chimeric genes were mixed and amplified with 3-XHO-EpoR-3066 and the 5' primers (5-RI-hKDR-1B, 5-RI-mKDR-1, and 5-RI-hTrkB-1) specific for each gene used in the previous cycle of amplification. The resulting PCR products were digested with EcoRI and XhoI and cloned into pcDNA3.1(+) (Invitrogen) to generate the plasmids phKE8 (human KDR/EpoR fusion), pmKE2 (flk-1/EpoR fusion), and phTE (TrkB/EpoR fusion).

[0252] Construction of cell lines for flow cytometry. CHO-K1 cells (American Type Culture Collection, Manassas, Va.) were stably transfected using Lipofectamine 2000 (Invitrogen) with either pcDNA 3.1 (Invitrogen) alone, pmKE2 alone, or a mixture of pcDNA 3.1 and a plasmid encoding full-length human KDR (Origene Inc., clone PR1371-H11). Stable transfectants were selected and maintained in the presence of 0.5 mg/ml of Geneticin (Invitrogen). The human KDR-expressing clone designated CHO-KDR and the murine VEGFR-2/EpoR-chimera-expressing population designated CHO-Flk were obtained by fluorescence activated cell sorting of the transfected population following staining with an anti-KDR polyclonal antiserum (R&D Systems). CHO-KDR and CHO-Flk cell lines were grown routinely in Dulbecco's modified Eagle's medium (DMEM; Gibco) supplemented with 10% (v/v) fetal bovine serum (FBS), 0.5 mg/ml Geneticin, 100 U/ml penicillin, 0.25 .mu.g/ml amphotericin B, 100 .mu.g/ml streptomycin and 2 mM L-glutamine.

[0253] Construction of Ba/F3 cell lines. Cell lines that would proliferate in response to VEGF binding by VEGFR-2 were constructed by transfection of the murine pre-B cell line Ba/F3 (DSMZ, Braunschweig, Germany) with phKE8 or pmKE2, receptor chimeras consisting of the extracellular domains of human or murine VEGFR-2 fused to the transmembrane and cytoplasmic domains of the human erythropoietin receptor (see above). Ba/F3 cells were maintained in minimal Ba/F3 medium (RPMI-1640 (Gibco) containing 10% FBS, 100 U/ml penicillin, 0.25 .mu.g/ml amphotericin B, 100 .mu.g/ml streptomycin and 2 mM L-glutamine) supplemented with 10% conditioned medium from WEHI-3B cells (DSMZ; grown in Iscove's modified Dulbecco's medium (Gibco)/10% FBS/25 .mu.M .beta.-mercaptoethanol) as a source of essential growth factors. Following electroporation with the plasmids pmKE2 or phKE8, stable transfectants were selected in 0.75 mg/ml Geneticin. Geneticin-resistant populations were transferred to minimal Ba/F3 medium containing 100 ng/ml of human VEGF.sub.165 (R&D Systems), and the resulting VEGF-dependent populations were designated Ba/F3-Flk and Ba/F3-KDR. Control cell line expressing a chimeric TrkB receptor (Ba/F3-TrkB) that would be responsive to stimulation by NT-4, the natural ligand for TrkB was similarly constructed using the plasmid phTE and human NT-4 (R&D Systems).

Analysis of Cell Surface Binding of Fibronectin-Based Scaffold Proteins:

[0254] Binding of fibronectin-based scaffold protein to cell-surface KDR and Flk-1 was analyzed simultaneously on VEGF-R2-expressing and control cells by flow cytometry. CHO-pcDNA3 cells (control) were released from their dishes with trypsin-EDTA, washed in Dulbecco's PBS without calcium and magnesium (D-PBS.sup.-; Invitrogen), and stained for 30 minutes at 37.degree. C. with 1.5 .mu.M CMTMR (5-(and-6)-(((4-chloromethyl)benzoyl)amino)-tetramethylrhodamine) (Molecular Probes). The cells were washed in D-PBS.sup.- and incubated for a further 30 minutes at 37.degree. C., and then resuspended in blocking buffer (D-PBS.sup.-/10% fetal bovine serum) on ice. CHO-KDR or CHO-Flk cells were treated identically except that CMTMR was omitted. 75,000 of CMTMR-stained CHO-pcDNA3 cells were mixed with an equal number of unstained CHO-KDR or CHO-Flk cells in each well of a V-bottom 96-well plate. All antibodies and fibronectin-based scaffold proteins were diluted in 25 .mu.l/well of blocking buffer, and each treatment was conducted for 1 hour at 4.degree. C. Cell mixtures were stained with His.sub.6-tagged fibronectin-based scaffold proteins, washed twice with cold D-PBS.sup.-, and then treated with 2.5 .mu.g/ml anti-His.sub.6 MAb (R&D Systems), washed, and stained with 4 .mu.g/ml Alexa Fluor 488-conjugated anti-mouse antibody (Molecular Probes). For cells treated with an anti-KDR mouse monoclonal antibody (Accurate Chemical, Westbury, N.Y.) or an anti-flk-1 goat polyclonal antibody (R&D Systems), the anti-His.sub.6 step was omitted, and antibody binding was detected with the species-appropriate Alexa Fluor 488 conjugated secondary antibody (Molecular Probes). Following staining, cells were resuspended in 200 .mu.l/well D-PBS.sup.-/1% FBS/1 .mu.g/ml 7-aminoactinomycin D (7-AAD; Molecular Probes) and analyzed by flow cytometry on a FACSCalibur (Becton Dickinson, San Jose, Calif.) equipped with a 488 nM laser. Following gating to exclude dead cells (7-AAD positive), VEGFR-2-expressing cells and CHO-pcDNA3 cells were measured independently for Alexa Fluor 488 fluorescence by gating on the CMTMR-negative or -positive populations, respectively. Control experiments showed that staining with CMTMR did not interfere with the detection of Alexa Fluor 488-conjugated antibodies on the surface of the stained cells.

[0255] Cell-surface binding was also assessed by fluorescence microscopy using the secondary antibodies described above. For these studies, antibodies were diluted in D-PBS containing calcium and magnesium (D-PBS.sup.+)/10% FBS. Cells were grown on 24- or 96-well plates, and following staining were kept in D-PBS.sup.+ for observation on an inverted fluorescence microscope.

Ba/F3 Cell Proliferation Assay:

[0256] Ba/F3 cells were washed three times in minimal Ba/F3 medium and resuspended in the same medium containing 15.8 ng/ml of proliferation factor (human VEGF.sub.165, murine VEGF.sub.164, or hNT-4 for Ba/F3-KDR, Ba/F3-Flk, or Ba/F3-TrkB cells, respectively), and 95 .mu.l containing 5.times.10.sup.4 Ba/F3-KDR cells or 2.times.10.sup.4 Ba/F3-Flk or Ba/F3-TrkB cells were added per well to a 96-well tissue culture plate. 5 .mu.l of serial dilutions of test protein in PBS was added to each well for a final volume of 100 .mu.l Ba/F3 medium/5% PBS/15 ng/ml growth factor. After incubation for 72 hours at 37.degree. C., proliferation was measured by addition of 20 .mu.l of CellTiter 96.RTM. Aqueous One Solution Reagent (Promega) to each well, incubation for 4 hours at 37.degree. C., and measurement of the absorbance at 490 nm using a microtiter plate reader (Molecular Dynamics).

HUVEC Cell Proliferation Assay:

[0257] HUVEC cells (Clonetics, Walkersville, Md.) from passage 2-6 were grown in EGM-2 medium (Clonetics). 5000 cells/well were resuspended in 200 .mu.l starvation medium (equal volumes of DMEM (Gibco) and F-12K medium (ATCC), supplemented with 0.2% fetal bovine serum and 1.times. penicillin/streptomycin/fungizone solution (Gibco)), plated in 96-well tissue culture plates and incubated for 48 hours. Fibronectin-based binding proteins were added to the wells and incubated for 1 hour at 37.degree., and then human VEGF.sub.165 was added to a final concentration of 16 ng/ml. After 48 hours incubation, cell viability was measured by addition of 30 .mu.l/well of a mixture of 1.9 mg/ml CellTiter96.RTM. AQueous MTS reagent (Promega) with 44 .mu.g/ml phenazine methosulfate (Sigma) and measurement of absorbance at 490 nm as described above for Ba/F3 cells.

Example 12

Antibody Light Chain-Based VEGFR Binding Polypeptides

[0258] FIGS. 21A and 21B show amino acid sequences of VEGFR binding polypeptides (SEQ ID NOs:241-310) based on an antibody light chain variable region (VL) framework/scaffold.

[0259] Light chain variable domain proteins were generated using the PROfusion.TM. system, as described above for use with .sup.10Fn3-derived proteins.

[0260] All references cited herein are hereby incorporated by reference in their entirety.

TABLE-US-00016 TABLE 1 Preferred Specific Peptide Sequences SEQ Binding Kd Binding Kd ID Clone N- DE to 1 nM KDR, to 1 nM FLK, NO Name terminus BC Loop Loop FG Loop KDR, % nM FLK, % nM KDR Binders 6 K1 Del 1-8 RHPHFPTR LQPPT M G L Y G H E L L T P 48 0.55 7 K2 Del 1-8 RHPHFPTR LQPPT D G E N G Q F L L V P 48 1.19 8 K5 Del 1-8 RHPHFPTR LQPPT M G P N D N E L L T P 47 1.54 9 K3 Del 1-8 RHPHFPTR LQPPT A G W D D H E L F I P 45 1.15 10 K7 Del 1-8 RHPHFPTR LQPPT S G H N D H M L M I P 40 2.2 11 K4 Del 1-8 RHPHFPTR LQPPT A G Y N D Q I L M T P 38 1.95 12 K9 Del 1-8 RHPHFPTR LQPPT F G L Y G K E L L I P 35 1.8 13 K10 Del 1-8 RHPHFPTR LQPPT T G P N D R L L F V P 33 0.57 14 K12 Del 1-8 RHPHFPTR LQPPT D V Y N D H E I K T P 29 0.62 15 K6 Del 1-8 RHPHFPTR LQPPT D G K D G R V L L T P 27 0.93 16 K15 Del 1-8 RHPHFPTR LQPPT E V H H D R E I K T P 25 0.35 17 K11 Del 1-8 RHPHFPTR LQPPT Q A P N D R V L Y T P 24 1.16 18 K14 Del 1-8 RHPHFPTR LQPPT R E E N D H E L L I P 20 0.57 19 K8 Del 1-8 RHPHFPTR LQPPT V T H N G H P L M T P 18 3.3 20 K13 Del 1-8 RHPHFPTR LQPPT L A L K G H E L L T P 17 0.58 21 VR28 WT RHPHFPTR LQPPT V A Q N D H E L I T P 3 11 22 159 WT RHPHFPTR LQPPA M A Q S G H E L F T P KDR and FLK Binders 24 E29 Del 1-8 RHPHFPTR LQPPT V E R N G R V L M T P 41 44 1.51 0.91 25 E19 Del 1-8 RHPHFPTR LQPPT V E R N G R H L M T P 38 40 1.3 0.66 33 E25 Del 1-8 RHPHFPTR LQPPT L E R N G R E L M T P 41 28 1.58 1.3 45 E9 Del 1-8 RHPHFPTR LQPPT E E R N G R T L R T P 24 34 2.37 1.4 50 E24 Del 1-8 RHPHFPTR LQPPT V E R N D R V L F T P 24 29 54 E26 Del 1-8 RHPHFPTR LQPPT V E R N G R E L M T P 27 20 1.66 2.05 59 E28 Del 1-8 RHPHFPTR LQPPT L E R N G R E L M V P 19 21 1.63 2.1 60 E3 Del 1-8 RHPHFPTR LQPPT D G R N D R K L M V P 37 14 0.96 5.4 65 E5 Del 1-8 RHPHFPTR LQPPT D G Q N G R L L N V P 26 10 0.4 3.2 91 E23 Del 1-8 RHHPHFPTR LQPPT V H W N G R E L M T P 36 7 92 E8 Del 1-8 RHPHFPTR LQPPT E E W N G R V L M T P 51 10 93 E27 Del 1-8 RHPHFPTR LQPPT V E R N G H T L M T P 37 9 94 E16 Del 1-8 RHPHFPTR LQPPT V E E N G R Q L M T P 35 0 95 E14 Del 1-8 RHPHFPTR LQPPT L E R N G Q V L F T P 33 11 96 E20 Del 1-8 RHPHFPTR LQPPT V E R N G Q V L Y T P 43 11 97 E21 Del 1-8 RHPHFPTR LQPPT W G Y K D H E L L I P 47 1 98 E22 Del 1-8 RHPHFPTR LQPPT L G R N D R E L L T P 45 3 99 E2 Del 1-8 RHPHFPTR LQPPT D G P N D R L L N I P 53 10 100 E12 Del 1-8 RHPHFPTR LQPPT F A R D G H E I L T P 36 1 101 E13 Del 1-8 RHPHFPTR LQPPT L E Q N G R E L M T P 38 1 102 E17 Del 1-8 RHPHFPTR LQPPT V E E N G R V L N T P 32 10 103 E15 Del 1-8 RHPHFPTR LQPPT L E P N G R Y L M V P 52 2 104 E10 Del 1-8 RHPHFPTR LQPPT E G R N G R E L F I P 53 3 154 M2 WT RHPHFPTR LQPPA W E R N G R E L F T P 156 M3 WT RHPHFPTR LQPPA K E R N G R E L F T P 172 M4 WT RHPHFPTH LQPPA T E R T G R E L F T P 173 M8 WT RHPHFPTH LQPPA K E R S G R E L F T P 175 M6 WT RHPHFPTH LQPPA L E R D G R E L F T P 180 M7 WT RHPHFPTR LQPTT W E R N G R E L F T P 181 M1 WT RHPHFPTR LQPTV L E R N D R E L F T P 177 M5FL WT RHPHFPTR LQPPL K E R N G R E L F T P

TABLE-US-00017 TABLE 2 KDR & FLK binders SEQ ID DE NO Clone Name N-terminus N-Terminus Framework 1 BC Loop Framework 2 Loop 23 D12 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 24 E29 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 25 E19 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 26 D1 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 27 C6 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 28 EGE5 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 29 EGE2 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 30 D4 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 31 E25 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 32 EGE6 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 33 C7 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 34 D9 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 35 EGE3 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 36 D3 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 37 D2 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 38 C8 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 39 EGE4 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 40 D7 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 41 D5 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 42 B3 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 43 E9 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 44 D6 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 45 EGE7 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 46 EGE1 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 47 F9 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 48 E24 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 49 B11 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 50 B12 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 51 B5 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 52 E26 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 53 C12 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 54 F4 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 55 E18 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 56 C11 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 57 E28 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 58 E3 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 59 F8 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 60 F3 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 61 B10 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 62 E6 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 63 E5 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 64 G4 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 65 A3 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 66 A4 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 67 A6 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 68 A7 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 69 A8 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 70 A9 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 71 A10 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 72 EGE11 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 73 A11 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 74 A12 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 75 B4 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 76 B6 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 77 B7, B8 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 78 B11 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 79 C1 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 80 C2 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 81 C3 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 82 C9 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 83 C10 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 84 D11 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 85 EGE8 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 86 EGE9 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 87 EGE10 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 88 EGE11 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 89 E23 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 90 E8 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 91 E27 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 92 E16 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 93 E14 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 94 E20 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 95 E21 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 96 E22 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 97 E2 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 98 E12 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 99 E13 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 100 E17 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 101 E15 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 102 E10 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 103 F1 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 104 F5 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 105 F6 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 106 F7 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 107 F10 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 108 F11 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 109 F12 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 110 G1 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 111 G2 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 112 G3 Del 1-8 EVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 113 MWF10 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 114 MWA10 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 115 MWA2 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 116 MWC10 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 117 MWB7 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 118 MWH8 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 119 MWA10 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 120 MWB2 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 121 MWC3-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 122 MWG11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 123 MWG11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 124 MWD3-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 125 MWE11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 126 MWD10 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 127 MWC1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 128 MWA12 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 129 MWB3-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 130 MWA11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 131 MWG12 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 132 MWH11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 133 MWD12 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 134 MWH5 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 135 MWA1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 136 MWG4-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 137 MWA12 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 138 MWG11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 139 MWC12 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 140 MWF11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 141 MWE11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 142 MWD10 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA

143 MWC4-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 144 MWF3 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 145 MWB2 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 146 MWE10 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 147 MWD9 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 148 MWH3-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 149 MWG10 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 150 MWH11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 151 MWF11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 152 M2 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 153 MWB09-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 154 M3 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPA 155 MWA3 WT VSDVPRDLEVVAATPTSLLISW LHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 156 MWE10 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 157 MWG3 WT VSDVPRDLEVVAATPTSLLISW LHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 158 MWD5 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 159 MWC3 WT VSDVPRDLEVVAATPTSLLISW LHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 160 MWH3 WT VSDVPRDLEVVAATPTSLLISW LHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 161 MWC2 WT VSDVPRDLEVVAATPTSLLISW LHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 162 MWE2 WT VSDVPRDLEVVAATPTSLLISW LHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 163 MWA2 WT VSDVPRDLEVVAATPTSLLISW FHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 164 MWD3 WT VSDVPRDLEVVAATPTSLLISW LHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 165 MWE3 WT VSDVPRDLEVVAATPTSLLISW LHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 166 MWB3 WT VSDVPRDLEVVAATPTSLLISW FHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 167 MWD2 WT VSDVPRDLEVVAATPTSLLISW LHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 168 MWC11 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 169 MWH12 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 170 M4 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 171 M8 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 172 MWF10-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 173 M6 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTH YYRITYGETGGNSPVQEFTVP LQPPA 174 MWB6 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPT 175 M5FL WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPL 176 MWG10-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPI 177 MWD08-f1; WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPI N42G 178 M7 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPTT 179 M1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP LQPTV 180 MWA07-f1 WT VSDVPRDLEVVAATPTSLLISW RPPHFPTR YYRITYGETGGNSPVQEFTVP LQPTV 181 MWH11-f1 WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP PQPPA 182 MWF09- WT VSDVPRDLEVVAATPTSLLISW RHPHFPTR YYRITYGETGGNSPVQEFTVP PQPPA f1;F48S 183 MWG12-f1 WT VSDVPRDLEVVAATPTSLLISW CHPHFPTR YYRITYGETGGNSPVQEFTVP LQPPI

TABLE-US-00018 SEQ Binding to Binding Kd Kd ID 1 nM to 1 nM KDR, Flk, NO Framework 3 FG Loop Framework 4 KDR, % Flk-1, % nM nM 23 ATISGLKPGVDYTITGYAVT V E R N G R K L M T P ISINYRT 46 47 24 ATISGLKPGVDYTITGYAVT V E R N G R V L M T P ISINYRT 41 44 1.51 0.91 25 ATISGLKPGVDYTITGYAVT V E R N G R H L M T P ISINYRT 38 40 1.3 0.66 26 ATISGLKPGVDYTITGYAVT V E R N G R M L M T P ISINYRT 38 38 27 ATISGLKPGVDYTITGYAVT L E R N G R V L M T P ISINYRT 36 49 28 ATISGLKPGVDYTITGYAVT L E R N G R V L N T P ISINYRT 32 47 29 ATISGLKPGVDYTITGYAVT V E R N G R Q L M T P ISINYRT 42 33 30 ATISGLKPGVDYTITGYAVT V E R N G R T L F T P ISINYRT 27 44 31 ATISGLKPGVDYTITGYAVT L E R N G R E L M T P ISINYRT 41 28 1.58 1.3 32 ATISGLKPGVDYTITGYAVT L E R N G R L L N T P ISINYRT 33 40 33 ATISGLKPGVDYTITGYAVT H E R N G R V L M T P ISINYRT 32 40 34 ATISGLRPGVDYTITGYAVT E E R N G R V L F T P ISINYRT 31 40 35 ATISGLKPGVDYTITGYAVT V E R N G R Q L Y T P ISINYRT 34 38 36 ATISGLKPGVDYTITGYAVT V E R N G R A L M T P ISINYRT 36 30 37 ATISGLKPGVDYTITGYAVT V E R N G R N L M T P ISINYRT 35 30 38 ATISGLKPGVDYTITGYAVT L E R N G R V L I T P ISINYRT 30 34 39 ATISGLKPGVDYTITGYAVT V E R N G R V L N T P ISINYRT 26 41 40 ATISGLKPGVDYTITGYAVT V E R N G K V L M T P ISINYRT 39 27 41 ATISGLKPGVDYTITGYAVT V E R N G R T L M M P ISINYRT 38 23 42 ATISGLKPGVDYTITGYAVT M E R N G R E L M T P ISINYRT 33 27 43 ATISGLKPGVDYTITGYAVT E E R N G R T L R T P ISINYRT 24 34 2.37 1.4 44 ATISGLKPGVDYTITGYAVT V E R N G K T L M T P ISINYRT 32 30 45 ATISGLKPGVDYTITGYAVT L E R N D R V L L T P ISINYRT 31 30 46 ATISGLKPGVDYTITGYAVT L E R N G R K L M T P ISINYRT 30 29 47 ATISGLKPGVDYTITGYAVT V E P N G R V L N T P ISINYRT 32 23 48 ATISGLKPGVDYTITGYAVT V E R N D R V L F T P ISINYRT 24 29 49 ATISGLKPGVDYTITGYAVT V E R N G R E L K T P ISINYRT 29 21 50 ATISGLKPGVDYTITGYAVT V E R N G R E L R T P ISINYRT 29 21 51 ATISGLKPGVDYTITGYAVT Q E R N G R E L M T P ISINYRT 27 24 52 ATISGLKPGVDYTITGYAVT V E R N G R E L M T P ISINYRT 27 20 1.66 2.05 53 ATISGLKPGVDYTITGYAVT V E R N G R V L S V P ISINYRT 24 20 54 ATISGLKPGVDYTITGYAVT V E R D G R T L R T P ISINYRT 31 18 55 ATISGLKPGVDYTITGYAVT V E R N G R E L N T P ISINYRT 17 29 1.2 0.53 56 ATISGLKPGVDYTITGYAVT V E R N G R V L I V P ISINYRT 19 21 57 ATISGLKPGVDYTITGYAVT L E R N G R E L M V P ISINYRT 19 21 1.63 2.1 58 ATISGLKPGVDYTITGYAVT D G R N D R K L M V P ISINYRT 37 14 0.96 5.4 59 ATISGLKPGVDYTITGYAVT V E H N G R T S F T P ISINYRT 33 13 60 ATISGLKPGVDYTITGYAVT V E R D G R K L Y T P ISINYRT 27 15 61 ATISGLKPGVDYTITGYAVT L E R N G R E L N T P ISINYRT 15 23 62 ATISGLKPGVDYTITGYAVT D G W N G R L L S I P ISINYRT 36 7 0.35 7.1 63 ATISGLKPGVDYTITGYAVT D G Q N G R L L N V P ISINYRT 26 10 0.4 3.2 64 ATISGLKPGVDYTITGYAVT I E K N G R H L N I P ISINYRT 21 12 65 ATISGLKPGVDYTITGYAVT D G W N G K M L S V P ISINYRT 33 7 66 ATISGLKPGVDYTITGYAVT D G Y N D R L L F I P ISINYRT 46 2 67 ATISGLKPGVDYTITGYAVT D G P N D R L L N I P ISINYRT 18 2 68 ATISGLKPGVDYTITGYAVT D G P N N R E L I V P ISINYRT 18 2 69 ATISGLKPGVDYTITGYAVT D G L N G K Y L F V P ISINYRT 38 4 70 ATISGLKPGVDYTITGYAVT E G W N D R E L F V P ISINYRT 31 4 71 ATISGLKPGVDYTITGYAVT F G W N G R E L L T P ISINYRT 34 4 72 ATISGLKPGVDYTITGYAVT F G W N D R E L L I P ISINYRT 50 0 73 ATISGLKPGVDYTITGYAVT L E W N N R V L M T P ISINYRT 26 6 74 ATISGLKPGVDYTITGYAVT V E W N G R V L M T P ISINYRT 40 10 75 ATISGLKPGVDYTITGYAVT N E R N G R E L M T P ISINYRT 19 12 76 ATISGLKPGVDYTITGYAVT L E R N G K E L M T P ISINYRT 23 11 77 ATISGLKPGVDYTITGYAVT V E R N G R E L L T P ISINYRT 18 10 78 ATISGLKPGVDYTITGYAVT V E R N G R E L K T P ISINYRT 29 21 79 ATISGLKPGVDYTITGYAVT Q E R N G R E L R T P ISINYRT 28 13 80 ATISGLKPGVDYTITGYAVT V E R N G R E L L W P ISINYRT 40 16 81 ATISGLKPGVDYTITGYAVT L E R N G R E L M I P ISINYRT 31 17 82 ATISGLKPGVDYTITGYAVT V E R N G L V L M T P ISINYRT 33 7 83 ATISGLKPGVDYTITGYAVT V E R N G R V L I I P ISINYRT 24 17 84 ATISGLKPGVDYTITGYAVT V E R N G H K L F T P ISINYRT 24 3 85 ATISGLKPGVDYTITGYAVT V E R N E R V L M T P ISINYRT 26 20 86 ATISGLKPGVDYTITGYAVT F G P N D R E L L T P ISINYRT 32 1 87 ATISGLKPGVDYTITGYAVT M G P N D R E L L T P ISINYRT 37 1 88 ATISGLKPGVDYTITGYAVT M G K N D R E L L T P ISINYRT 32 1 89 ATISGLKPGVDYTITGYAVT V H W N G R E L M T P ISINYRT 36 7 90 ATISGLKPGVDYTITGYAVT E E W N G R V L M T P ISINYRT 51 10 91 ATISGLKPGVDYTITGYAVT V E R N G H T L M T P ISINYRT 37 9 92 ATISGLKPGVDYTITGYAVT V E E N G R Q L M T P ISINYRT 35 0 93 ATISGLKPGVDYTITGYAVT L E R N G Q V L F T P ISINYRT 33 11 94 ATISGLKPGVDYTITGYAVT V E R N G Q V L Y T P ISINYRT 43 11 95 ATISGLKPGVDYTITGYAVT W G Y K D H E L L I P ISINYRT 47 1 96 ATISGLKPGVDYTITGYAVT L G R N D R E L L T P ISINYRT 45 3 97 ATISGLKPGVDYTITGYAVT D G P N D R L L N I P ISINYRT 53 10 98 ATISGLKPGVDYTITGYAVT F A R D G H E I L T P ISINYRT 36 1 99 ATISGLKPGVDYTITGYAVT L E Q N G R E L M T P ISINYRT 38 1 100 ATISGLKPGVDYTITGYAVT V E E N G R V L N T P ISINYRT 32 10 101 ATISGLKPGVDYTITGYAVT L E P N G R Y L M V P ISINYRT 52 2 102 ATISGLKPGVDYTITGYAVT E G R N G R E L F I P ISINYRT 53 3 103 ATISGLKPGVDYTITGYAVT S G R N D R E L L V P ISINYRT 18 2 104 ATISGLRPGVDYTITGYAVT V E R D G R E L N I P ISINYRT 12 8 105 ATISGLKPGVDYTITGYAVT V E Q N G R V L M T P ISTNYRT 37 2 106 ATISGLKPGVDYTITGYAVT V E H N G R V L N I P ISINYRT 30 7 107 ATISGLKPGVDYTITGYAVT M A P N G R E L L T P ISINYRT 29 1 108 ATISGLKPGVDYTITGYAVT V E Q N G R V L N T P ISINYRT 20 8 109 ATISGLKPGVDYTITGYAVT D G R N G H E L M T P ISINYRT 17 1 110 ATISGLKPGVDYTITGYAVT E G R N G R E L M V P ISINYRT 22 2 111 ATISGLKPGVDYTITGYAVT L E R N N R E L L T P ISiNYRT 25 9 112 ATISGLKPGVDYTITGYAVT M E R S G R E L M T P ISINYRT 28 10 113 ATISGLKPGVDYTITGYAVT R A L L S I E L F T P ISINYRT 114 ATISGLKPGVDYTITGYAVT F A R K G T E L F T P ISINYRT 115 ATISGLKPGVDYTITGYAVT L E R C G R E L F T P ISINYRT 116 ATISGLKPGVDYTITGYAVT R E R N G R E L F T P ISINYRT 117 ATISGLKPGVDYTITGYAVT K E R N G R E L F T P ISINYRT 118 ATISGLKPGVDYTITGYAVT C E R N G R E L F T P ISINYRT 119 ATISGLKPGVDYTITGYAVT L E R T G R E L F T P ISTNYRT 120 ATISGLKPGVDYTITGYAVT W E R T G K E L F T P ISINYRT 121 ATISGLKPGVDYTITGYAVT I E R T C R E L F T P ISINYRT 122 ATISGLKPGVDYTITGYAVT G G M I V R E L F T P ISINYRT 123 ATISGLKPGVDYTITGYAVT F G R S S R E L F T P ISINYRT 124 ATISGLKPGVDYTITGYAVT R H K S R G E L F T P ISINYRT 125 ATISGLKPGVDYTITGYAVT R H R D K R E L F T P ISINYRT 126 ATISGLKPGVDYTITGYAVT Y H R G R G E L F T P ISINYRT 127 ATISGLKPGVDYTITGYAVT R H R G C R E L F T P ISINYRT 128 ATISGLKPGVDYTITGYAVT S H R L R K E L F T P ISINYRT 129 ATISGLKPGVDYTITGYAVT M H R Q R G E L F T P ISINYRT 130 ATISGLKPGVDYTITGYAVT F H R R R G E L F T P ISINYRT 131 ATISGLKPGVDYTITGYAVT F H R R R G E L F T P ISINYRT 132 ATISGLKPGVDYTITGYAVT S H R R R N E L F T P ISTNYRT 133 ATISGLKPGVDYTITGYAVT L H R R V R E L F T P ISTNYRT 134 ATISGLKPGVDYTITGYAVT R H R R R G E L F T P ISINYRT 135 ATISGLKPGVDYTITGYAVT W H R S R K E L F T P ISINYRT 136 ATISGLKPGVDYTITGYAVT R H R S R G E L F T P ISINYRT 137 ATISGLKPGVDYTITGYAVT V H R T G R E L F T P ISINYRT 138 ATISGLKPGVDYTITGYAVT W H R V R G E L F T P ISINYRT 139 ATISGLKPGVDYTITGYAVT W H R V R G E L F T P ISINYRT 140 ATISGLKPGVDYTITGYAVT W H R W R G E L F T P ISTNYRT 141 ATISGLKPGVDYTITGYAVT W K R S G G E L F T P ISINYRT 142 ATISGLKPGVDYTITGYAVT R L X N X V E L F T P ISINYRT 143 ATISGLKPGVDYTITGYAVT W R T P H A E L F T P ISINYRT 144 ATISGLKPGVDYTITGYAVT L S P H S V E L F T P ISINYRT

145 ATISGLKPGVDYTITGYAVT V S R Q K A E L F T P ISINYRT 146 ATISGLKPGVDYTITGYAVT S S Y S K L E L F T P ISINYRT 147 ATISGLKPGVDYTITGYAVT L T D R G S E L F T P ISINYRT 148 ATISGLKPGVDYTITGYAVT G T R T R S E L F T P ISINYRT 149 ATISGLKPGVDYTITGYAVT P V A G C S E L F T P ISINYRT 150 ATISGLKPGVDYTITGYAVT W W Q T P R E L F T P ISINYRT 151 ATISGLKPGVDYTITGYAVT W W Q T P R E L F T P ISINYRT 152 ATISGLKPGVDYTITGYAVT W E R N G R E L F T P ISINYRT 153 ATISGLKPGVDYTITGYAVT W E W N G R E L F T P ISINYRT 154 ATISGLKPGVDYTITGYAVT K E R N G R E L F T P ISINYRT 155 ATISGLKPGVDYTITGYAVT G A L N T S E L F T P ISINYRT 156 ATISGLKPGVDYTITGYAVT F G R E R R E L F T P ISINYRT 157 ATISGLKPGVDYTITGYAVT S G R V S F E L F T P ISTNYRT 158 ATISGLKPGVDYTITGYAVT F H R R R G E L F T P ISINYRT 159 ATISGLKPGVDYTITGYAVT L I R M N T E L F T P ISINYRT 160 ATISGLKPGVDYTITGYAVT C L H L I T E L F T P ISINYRT 161 ATISGLKPGVDYTITGYAVT V L K L T L E L F T P ISINYRT 162 ATISGLKPGVDYTITGYAVT V L K L T L E L F T P ISINYRT 163 ATISGLKPGVDYTITGYAVT V L K L T L E L F T P ISINYRT 164 ATISGLKPGVDYTITGYAVT A L M A S G E L F T P ISINYRT 165 ATISGLKPGVDYTITGYAVT S M K N R L E L F T P ISINYRT 166 ATISGLKPGVDYTITGYAVT L R C L I P E L F T P ISINYRT 167 ATISGLKPGVDYTITGYAVT V S R Q K A E L F T P ISINYRT 168 ATISGLKPGVDYTITGYAVT W S R T G R E L F T P ISINYRT 169 ATISGLKPGVDYTITGYAVT V W R T G R E L F T P ISTNYRT 170 ATISGLKPGVDYTITGYAVT T E R T G R E L F T P ISINYRT 171 ATISGLKPGVDYTITGYAVT K E R S G R E L F T P ISINYRT 172 ATISGLKPGVDYTITGYAVT L E R N D R E L F T P ISINYRT 173 ATISGLKPGVDYTITGYAVT L E R D G R E L F T P ISINYRT 174 ATISGLKPGVDYTITGYAVT Q G R H K R E L F T P ISINYRT 175 ATISGLKPGVDYTITGYAVT K E R N G R E L F T P ISINYRT 176 ATISGLKPGVDYTITGYAVT M A Q N D H E L I T P ISINYRT 177 ATISGLKPGVDYTITGYAVT M A Q N D H E L I T P ISINYRT 178 ATISGLKPGVDYTITGYAVT W E R N G R E L F T P ISINYRT 179 ATISGLKPGVDYTITGYAVT L E R N D R E L F T P ISINYRT 180 ATISGLKPGVDYTITGYAVT L E R N D R E L F T P ISINYRT 181 ATISGLKPGVDYTITGYAVT K E R S G R E L F T P ISINYRT 182 ATISGLKPGVDYTITGYAVT L E R N D R E L F T P ISINYRT 183 ATISGLKPGVDYTITGYAVT M A Q N D H E L I T P ISINYRT

TABLE-US-00019 TABLE 3 KDR binders Binding to Kd SEQ ID 1 nM KDR, NO Clone Name N-terminus BC Loop DE Loop FG Loop KDR, % nM 6 K1 Del 1-8 RHPHFPTR LQPPT M G L Y G H E L L T P 48 0.55 7 K2 Del 1-8 RHPHFPTR LQPPT D G E N G Q F L L V P 48 1.19 8 K5 Del 1-8 RHPHFPTR LQPPT M G P N D N E L L T P 47 1.54 9 K3 Del 1-8 RHPHFPTR LQPPT A G W D D H E L F I P 45 1.15 311 3'E9 PR4 Del 1-8 RHPHFPTR LQPPT V E Q D G H V L Y I P 44 312 2'Del E6 PR4 Del 1-8 RHPHFPTR LQPPT M G K N G H E L L T P 43 313 3'D3 PR4 Del 1-8 RHPHFPTR LQPPT P G P G D R E L I T P 42 314 2'Del F8 PR4 Del 1-8 RHPHFPTR LQPPT A G P G A H E L L T P 42 315 4'B3 PR4 Del 1-8 RHPHFPTR LQPPT M A Q N N R E L L T P 42 316 3'E3 PR4 Del 1-8 RHPHFPTR LQPPT M A Q Y G R E L L T P 41 10 K7 Del 1-8 RHPHFPTR LQPPT S G H N D H M L M I P 40 2.2 317 3'H11 PR4 Del 1-8 RHPHFPTR LQPPT L A H N G N E L L T P 39 318 3'B4 PR4 Del 1-8 RHPHFPTR LQPPT V A W N G H E L M T P 38 11 K4 Del 1-8 RHPHFPTR LQPPT A G Y N D Q I L M T P 38 1.95 319 2'Del F7 PR4 Del 1-8 RHPHFPTR LQPPT L G L R D R E L F V P 38 320 2'Del D3 PR4 Del 1-8 RHPHFPTR LQPPT S G L N D R V L F I P 38 321 3'C6 PR4 Del 1-8 RHPHFPTR LQPPT M G P N D R E L L T P 37 322 3'F3 PR4 Del 1-8 RHPHFPTR LQPPT L G H N D R E L L T P 37 323 3'H3 PR4 Del 1-8 RHPHFPTR LQPPT L G L N D R E L M T P 36 324 1'Del G10 PR4 Del 1-8 RHPHFPTR LQPPT M A Q N G H K L M T P 36 12 K9 Del 1-8 RHPHFPTR LQPPT F G L Y G K E L L I P 35 1.8 325 2'DelE4 PR4 Del 1-8 RHPHFPTR LQPPT V H W N G H E L M T P 34 326 2'Del C6 PR4 Del 1-8 RHPHFPTR LQPPT M G F M A H E L M V P 34 327 2'Del C11 PR4 Del 1-8 RHPHFPTR LQPPT A G L N E H E L L I P 34 328 2'Del D10 PR4 Del 1-8 RHPHFPTR LQPPT L A D N A R E L L T P 34 329 2'Del H5 PR4 Del 1-8 RHPHFPTR LQPPT L G K D V R E L L T P 34 330 3'A7 PR4 Del 1-8 RHPHFPTR LQPPT L S D S G H A L F T P 34 331 2'Del E3 PR4 Del 1-8 RHPHFPTR LQPPT L G P Y E H E L L T P 33 13 K10 Del 1-8 RHPHFPTR LQPPT T G P N D R L L F V P 33 0.57 332 2'Del B5 PR4 Del 1-8 RHPHFPTR LQPPT A G R H D H E L I I P 33 333 3'C12 PR4 Del 1-8 RHPHFPTR LQPPT I G P N N H E L L T P 33 334 2'Del G9 PR4 Del 1-8 RHPHFPTR LQPPT V E Q N G R E L I I P 33 335 2'Del C1 PR4 Del 1-8 RHPHFPTR LQPPT A G L D E H E L L I P 32 336 3'E1 PR4 Del 1-8 RHPHFPTR LQPPT V A P N G H E L F T P 32 337 3'C3 PR4 Del 1-8 RHPHFPTR LQPPT M A Q N G H A L F T P 32 338 2'DelB7 PR4 Del 1-8 RHPHFPTR LQPPT V G Y N N R E L L T P 32 339 3'F1 PR4 Del 1-8 RHPHFPTR LQPPT V A Q D G H F L Y T P 31 340 2'Del B4 PR4 Del 1-8 RHPHFPTR LQPPT S G H N G H E V M T P 31 341 3'G3 PR4 Del 1-8 RHPHFPTR LQPPT F D Q S D H E L L T P 31 342 2'DelH4 PR4 Del 1-8 RHPHFPTR LQPPT V G P N E R M L M T P 30 343 3'D9 PR4 Del 1-8 RHPHFPTR LQPPT G Y Y N D R E L L T P 30 344 3'G10 PR4 Del 1-8 RHPHFPTR LQPPT L T H N D H E L L T P 30 345 3'B2 PR4 Del 1-8 RHPHFPTR LQPPT V G R N D R E L L T P 29 346 2'DelC3 PR4 Del 1-8 RHPHFPTR LQPPT W A Q N G R E L L T P 29 347 3'F2 PR4 Del 1-8 RHPHFPTR LQPPT L G K N D H E L L T P 29 348 4'C9 PR4 Del 1-8 RHPHFPTR LQPPT L G P N D H E L M T P 29 349 2'Del B2 PR4 Del 1-8 RHPHFPTR LQPPT T G W N G N E L F T P 29 14 K12 Del 1-8 RHPHFPTR LQPPT D V Y N D H E I K T P 29 0.62 350 4'H7 PR4 Del 1-8 RHPHFPTR LQPPT L A H N D H E L L T P 29 351 2'Del D1 PR4 Del 1-8 RHPHFPTR LQPPT L E Q N D R V L L T P 28 352 2'Del H6 PR4 Del 1-8 RHPHFPTR LQPPT T G H H D H E L I I P 28 353 3'B12 PR4 Del 1-8 RHPHFPTR LQPPT V A H E N R E L L T P 28 354 4'C5 PR4 Del 1-8 RHPHFPTR LQPPT L G L N D H E L I T P 27 15 K6 Del 1-8 RHPHFPTR LQPPT D G K D G R V L L T P 27 0.93 355 3'D8 PR4 Del 1-8 RHPHFPTR LQPPT A G P N D H Q L F T P 27 356 3'C5 PR4 Del 1-8 RHPHFPTR LQPPT D A M Y G R E L M T P 27 357 3'A8 PR4 Del 1-8 RHPHFPTR LQPPT V A W D D H E L L T P 27 358 2'Del F11 PR4 Del 1-8 RHPHFPTR LQPPT M G Q N D K E L I T P 27 359 4'D8 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N G H E L Y T P 26 360 2'Del C5 PR4 Del 1-8 RHPHFPTR LQPPT P G H N D H E L M V P 26 16 K15 Del 1-8 RHPHFPTR LQPPT E V H H D R E I K T P 25 0.35 361 3'B1 PR4 Del 1-8 RHPHFPTR LQPPT E A R N G R E L L T P 25 362 3'A9 PR4 Del 1-8 RHPHFPTR LQPPT L A H N D R E L L T P 25 363 4'B11 PR4 Del 1-8 RHPHFPTR LQPPT M A H N D H E L L T P 25 17 K11 Del 1-8 RHPHFPTR LQPPT Q A P N D R V L Y T P 24 1.16 364 3D12 PR3 Del 1-8 RHPHFPTR LQPPT L G Q N D R Q L L V P 24 365 2'Del H12 PR4 Del 1-8 RHPHFPTR LQPPT A G G N G H E L L T P 24 366 3'H9 PR4 Del 1-8 RHPHFPTR LQPPT H G P Y D Q V L L T P 24 367 3'F6 PR4 Del 1-8 RHPHFPTR LQPPT I E Q S G L Q L M T P 24 368 1'DelE6 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N D R E L L T P 24 369 3'E5 PR4 Del 1-8 RHPHFPTR LQPPT V A W D G R E L F T P 23 370 3A3 PR3 Del 1-8 RHPHFPTR LQPPT L A Y N G R E I I T P 23 371 3A2 PR3 Del 1-8 RHPHFPTR LQPPT W S Q N N R E L F T P 23 372 3'B11 PR4 Del 1-8 RHPHFPTR LQPPT E T W N D H E I R T P 23 373 1'DelA2 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N G H Q L F T P 23 374 2'D6-PR4 WT RHPHFPTR LQPPT V T H N G H P L M T P 22 375 3'H1 PR4 Del 1-8 RHPHFPTR LQPPT F A Q N D H Q L F T P 22 376 2'Del G11 PR4 Del 1-8 RHPHFPTR LQPPT G G Q M N R V L M T P 22 377 2'Del F5 PR4 Del 1-8 RHPHFPTR LQPPT L V H N D R E L L T P 22 378 1'DelE7 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N G H E L F T P 22 379 2'E4-PR4 WT RHPHFPTR LQPPT V H W N G H E L M T P 22 380 2'Del F6 PR4 Del 1-8 RHPHFPTR LQPPT L G W N D H E L Y I P 22 381 3'E10 PR4 Del 1-8 RHPHFPTR LQPPT A G H K D Q E L L T P 21 382 4'A9 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N N H E L L T P 21 383 4'G12 PR4 Del 1-8 RHPHFPTR LQPPT V A W N D H E I Y T P 21 384 3'B10 PR4 Del 1-8 RHPHFPTR LQPPT L A Q T G R E L L T P 21 385 2'DelH9 PR4 Del 1-8 RHPHFPTR LQPPT V G W S G H E L F V P 20 386 3'H8 PR4 Del 1-8 RHPHFPTR LQPPT V G H N D R E L I T P 20 387 2'DelA5 PR4 Del 1-8 RHPHFPTR LQPPT W N Q N G Q E L F T P 20 388 3B5 PR3 Del 1-8 RHPHFPTR LQPPT F G Q N G H A L L T P 20 389 3C7 PR3 Del 1-8 RHPHFPTR LQPPT R G L N D G E L L T P 20 390 3G2 PR3 Del 1-8 RHPHFPTR LQPPT F G P S D H V L L T P 20 18 K14 Del 1-8 RHPHFPTR LQPPT R E E N D H E L L I P 20 0.57 391 4'B12 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N N H E L L T P 20 392 4'B8 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N D H K L F I P 20 393 2'Del F1 PR4 Del 1-8 RHPHFPTR LQPPT R D Q Y E H E L L T P 20 394 3'G1 PR4 Del 1-8 RHPHFPTR LQPPT L A L N G H E L F T P 19 395 3'D2 PR4 Del 1-8 RHPHFPTR LQPPT V E S N G H A L F V P 19 396 2'DelG5 PR4 Del 1-8 RHPHFPTR LQPPT V G Q N N H E L L T P 19 397 2'DelC7 PR4 Del 1-8 RHPHFPTR LQPPT W D Q N G H V L L T P 19 398 2'Del E5 PR4 Del 1-8 RHPHFPTR LQPPT E G L N D H E L I I P 19 399 3'C8 PR4 Del 1-8 RHPHFPTR LQPPT E G L N D H E L M I P 19 400 3'G7 PR4 Del 1-8 RHPHFPTR LQPPT E G Q N D Q L L F I P 19 401 3'A6 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N G H E L L T P 19 402 4'G4 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N D R E L L T P 19 403 4'H5 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N G H E L F T P 18 404 1'DelH12 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N E R E L F T P 18 19 K8 Del 1-8 RHPHFPTR LQPPT V T H N G H P L M T P 18 3.3 405 2'Del D5 PR4 Del 1-8 RHPHFPTR LQPPT V A W N D H M L M T P 18 406 3'F9 PR4 Del 1-8 RHPHFPTR LQPPT L G P N D R E L M T P 18 407 2'H4-PR4 WT RHPHFPTR LQPPT V G P N E R M L M T P 17 408 4'H12 PR4 Del 1-8 RHPHFPTR LQPPT V A H N D H E L L T P 17 409 1'DelD2 PR4 Del 1-8 RHPHFPTR LQPPT V A K N D H E L L T P 17 410 4'E7 PR4 Del 1-8 RHPHFPTR LQPPT W A Q N D H E L L T P 17 411 1'DelH10 PR4 Del 1-8 RHPHFPTR LQPPT F A Q N D H E L L T P 17 20 K13 Del 1-8 RHPHFPTR LQPPT L A L K G H E L L T P 17 0.58 412 3C3 PR3 Del 1-8 RHPHFPTR LQPPT M E Q N G H E L F T P 17 413 2'Del B3 PR4 Del 1-8 RHPHFPTR LQPPT D A P N G R E L M V P 17 414 2'Del A2 PR4 Del 1-8 RHPHFPTR LQPPT G G R N G H T L L T P 17 415 3'F12 PR4 Del 1-8 RHPHFPTR LQPPT L S Q T D H E L L T P 17 416 3B4 PR3 Del 1-8 RHPHFPTR LQPPT V G Q N E H E L L T P 17 417 3F8 PR3 Del 1-8 RHPHFPTR LQPPT V A Q N G H E L K T P 17 418 1'DelH5 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N D R E L F T P 17 419 1'DelD5 PR4 Del 1-8 RHPHFPTR LQPPT V G Q N H H E L F T P 17 420 3'E11 PR4 Del 1-8 RHPHFPTR LQPPT V G P H D R E L L T P 17 421 2'C6-PR4 WT RHPHFPTR LQPPT M G F M A H E L M V P 16 422 4C9 PR3 Del 1-8 RHPHFPTR LQPPT L A Q N D H E L L T P 16 423 3C9 PR3 Del 1-8 RHPHFPTR LQPPT L V R N D H E L L T P 16 424 3F10 PR3 Del 1-8 RHPHFPTR LQPPT L A Q D D H E L L T P 16 425 2'Del A11 PR4 Del 1-8 RHPHFPTR LQPPT E D I R V L W L N T T 16 426 1'DelD1 PR4 Del 1-8 RHPHFPTR LQPPT V T Q N D H E L L T P 16 427 1'DelE2 PR4 Del 1-8 RHPHFPTR LQPPT V G Q N D H E L L T P 16 428 1'DelF3 PR4 Del 1-8 RHPHFPTR LQPPT M A Q N D H K L F T P 16 429 4'A5 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N D H E L L T P 16 430 1'DelB8 PR4 Del 1-8 RHPHFPTR LQPPT M A Q N D H E L L T P 16 431 4'B7 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N N H E L L T P 16 432 4F4 PR3 Del 1-8 RHPHFPTR LQPPT L A Q N D R E L I T P 15 433 4B11 PR3 Del 1-8 RHPHFPTR LQPPT V G Q N N H E L I T P 15 434 3'G2 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N G H E L I T P 15 435 2'Del C8 PR4 Del 1-8 RHPHFPTR LQPPT T A H N G H E L L T P 15 436 3'B8 PR4 Del 1-8 RHPHFPTR LQPPT L G Y H D H A L F T P 14 437 3'H10 PR4 Del 1-8 RHPHFPTR LQPPT W A W N D H E L M T P 14 438 1'DelA1 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N D H E L L T P 14 439 4'D6 PR4 Del 1-8 RHPHFPTR LQPPT M A Q N D H E L M T P 14 440 4F9 PR3 Del 1-8 RHPHFPTR LQPPT M A Q N D H E L L T P 14 441 4H5 PR3 Del 1-8 RHPHFPTR LQPPT V A Q N G H E L I T P 14 442 2D12 PR3 WT RHPHFPTR LQPPT E G W I D H E I M I P 14 443 3'F7 PR4 Del 1-8 RHPHFPTR LQPPT E G Q N G S E L I V P 14 444 4C11 PR3 Del 1-8 RHPHFPTR LQPPT M A Q N D R E L I T P 14 445 4B6 PR3 Del 1-8 RHPHFPTR LQPPT V G Q N D H E L F T P 14 446 1'DelE12 PR4 Del 1-8 RHPHFPTR LQPPT V A Q S D H E L F T P 13 447 1'DelC2 PR4 Del 1-8 RHPHFPTR LQPPT V D R N D H E L F T P 13 448 1'DelA9 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N D H E L M T P 13 449 1'DelA4 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N D H E L F T P 13 450 3G5 PR3 Del 1-8 RHPHFPTR LQPPT L G E N D R K L I T P 13 451 4A12 PR3 Del 1-8 RHPHFPTR LQPPT V A Q N D H E L L T P 13 452 2'Del E12 PR4 Del 1-8 RHPHFPTR LQPPT E G P N G H E L I T P 13 453 3G1 PR3 Del 1-8 RHPHFPTR LQPPT M A Q N V R E L L T P 13 454 4F12 PR3 Del 1-8 RHPHFPTR LQPPT V T Q N G H E L I T P 13 455 4B7 PR3 Del 1-8 RHPHFPTR LQPPT V T Q N D H E L F T P 13 456 4'G8 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N G H E L L T P 13 457 3'E8 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N D R Q L F T P 12 458 3'E4 PR4 Del 1-8 RHPHFPTR LQPPT V G P N D R E L I V P 12 459 1'DelC6 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N E H E L L T P 12 460 1'DelD3 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N N H E L I T P 12 461 3A8 PR3 Del 1-8 RHPHFPTR LQPPT E A H H G H E L M I P 12 462 3C5 PR3 Del 1-8 RHPHFPTR LQPPT G D H N D R E L M T P 12 463 2'G11-PR4 WT RHPHFPTR LQPPT G G Q M N R V L M T P 12 464 3'D4 PR4 Del 1-8 RHPHFPTR LQPPT L A H N D R E L I T P 12 465 3E6 PR3 Del 1-8 RHPHFPTR LQPPT V P Q N G H E L I T M 12 466 1'DelA11 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N D H E L F T P 12 467 4'D12 PR4 Del 1-8 RHPHFPTR LQPPT V D Q N D H E L F T P 12 468 2'D5-PR4 WT RHPHFPTR LQPPT V A W N D H M L M T P 11 469 2'A1-PR4 WT RHPHFPTR LQPPT S G H N D H M L M I P 11 470 1'DelG11 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N G H V L I T P 11 471 2'DelB10 PR4 Del 1-8 RHPHFPTR LQPPT V T H N D H E L L T P 11 472 2'DelB11 PR4 Del 1-8 RHPHFPTR LQPPT V G Q N D H E L M T P 11 473 1'DelC5 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N D H E I M T P 11 474 4'B6 PR4 Del 1-8 RHPHFPTR LQPPT L A Q N D H E L I T P 11 475 3H9 PR3 Del 1-8 RHPHFPTR LQPPT V S Q Q N H E L L T P 11 476 4E10 PR3 Del 1-8 RHPHFPTR LQPPT V A Q N D H E L M T P 11 477 3F5 PR3 Del 1-8 RHPHFPTR LQPPT V A Y N E H E L Y T P 11 478 4A9 PR3 Del 1-8 RHPHFPTR LQPPT V A Q H D H E L L T P 11 479 1'DelH7 PR4 Del 1-8 RHPHFPTR LQPPT V G Q N D Q E L L T P 11 480 1'DelB10 PR4 Del 1-8 RHPHFPTR LQPPT V A R N D H E L M T P 11 481 2'DelB9 PR4 Del 1-8 RHPHFPTR LQPPT V G P T D H E L L T P 11 482 3F11 PR3 Del 1-8 RHPHFPTR LQPPT V G L T D H V L L T P 10 483 4C4 PR3 Del 1-8 RHPHFPTR LQPPT V A Q D D H E L F T P 10 484 4B5 PR3 Del 1-8 RHPHFPTR LQPPT L A Q N D H E L F T P 10 485 3D4 PR3 Del 1-8 RHPHFPTR LQPPT V G W N D H E L I T P 10 486 4A4 PR3 Del 1-8 RHPHFPTR LQPPT V A Q N D H E L F T P 10 487 3D11 PR3 Del 1-8 RHPHFPTR LQPPT L G Q E N Q E L I T P 10 488 2H10 PR3 WT RHPHFPTR LQPPT L A P S A R E L M T P 10 489 3G10 PR3 Del 1-8 RHPHFPTR LQPPT V V H N G H E I L T P 10 490 3F4 PR3 Del 1-8 RHPHFPTR LQPPT M G Y E D H E L I T P 10 491 2H12 PR3 WT RHPHFPTR LQPPT E G Y Q N H E L S V P 10 492 4C2 PR3 Del 1-8 RHPHFPTR LQPPT V D Q N D H E L F T P 10 493 1'DelG9 PR4 Del 1-8 RHPHFPTR LQPPT V A Q S D H E L M T P 10 494 1'DelH9 PR4 Del 1-8 RHPHFPTR LQPPT V G Q N D H E L I T P 10 495 1'DelB3 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N D H E L M T P 10 496 1'DelH1 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N G H E L I T P 9 497 3'A3 PR4 Del 1-8 RHPHFPTR LQPPT R A Q N D H E L I T P 9 498 1'DelC4 PR4 Del 1-8 RHPHFPTR LQPPT V A Q S N H E L M T P 9 499 1'DelE11 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N D R E L I T P 9 500 3F1 PR3 Del 1-8 RHPHFPTR LQPPT L T H N E Q Y L F T P 9 501 2G9 PR3 WT RHPHFPTR LQPPT E I Y N D H E L M T P 9 502 3'D11 PR4 Del 1-8 RHPHFPTR LQPPT M A Q N D H E L I T P 9 503 2'DelH2 PR4 Del 1-8 RHPHFPTR LQPPT V S Q Y G H E L I T P 8 504 1'DelC10 PR4 Del 1-8 RHPHFPTR LQPPT V A K N D H E L I T P 8 505 4D2 PR3 Del 1-8 RHPHFPTR LQPPT V A Q N N H E L I T P 8 506 4A9 PR3 Del 1-8 RHPHFPTR LQPPT V A Q H D H E L L T P 8 507 2F3 PR3 WT RHPHFPTR LQPPT L S H Y G K E L R T P 8 508 4A2 PR3 Del 1-8 RHPHFPTR LQPPT V A Q N A H E L M T P 8 509 4G4 PR3 Del 1-8 RHPHFPTR LQPPT L G Q N D H E L L T P 8 510 1'DelB7 PR4 Del 1-8 RHPHFPTR LQPPT V A Q N D H E L K T P 8 511 3'D12 PR4 Del 1-8 RHPHFPTR LQPPT G E Q N D Y E L L V P 7 512 2'Del F12 PR4 Del 1-8 RHPHFPTR LQPPT L T Q H D H E L L T P 7 513 4E2 PR3 Del 1-8 RHPHFPTR LQPPT M A Q N D H E L I T P 7 514 2C9 PR3 WT RHPHFPTR LQPPT E A P N G R E L R T P 7 515 2'B9-PR4 WT RHPHFPTR LQPPT V G P T D H E L L T P 7 516 1'DelH6 PR4 Del 1-8 RHPHFPTR LQPPT V G Q Y D H E L I T P 6 517 4A3 PR3 Del 1-8 RHPHFPTR LQPPT V A Q D E H E L I T P 6 518 2C12 PR3 WT RHPHFPTR LQPPT D A Q N V Q A P I A Q 6 519 2G12 PR3 WT RHPHFPTR LQPPT S G Q N D H A L L I P 6 520 2'A11-PR4 WT RHPHFPTR LQPPT E D I R V L W L N T T 6 521 2'C7-PR4 WT RHPHFPTR LQPPT W D Q N G H V L L T P 5 522 2'F7-PR4 WT RHPHFPTR LQPPT L G L R D R E L F V P 5 523 2C6 PR3 WT RHPHFPTR LQPPT V E P N G H K L A I P 5 524 3'E6 PR4 Del 1-8 RHPHFPTR LQPPT F G Q N G K E F R I P 5 525 2'B4-PR4 WT RHPHFPTR LQPPT S G H N G H E V M T P 4 526 2'F6-PR4 WT RHPHFPTR LQPPT L G W N D H E L Y I P 4 527 2'H5-PR4 WT RHPHFPTR LQPPT L G K D V R E L L T P 3 528 2F10 PR3 WT RHPHFPTR LQPPT L A L F D H E L L T P 3 21 VR28 WT RHPHFPTR LQPPT V A Q N D H E L I T P 3 11 22 159 WT RHPHFPTR LQPPA M A Q S G H E L F T P

TABLE-US-00020 TABLE 4 Sequences of characterized VEGF-R2 binding clones ##STR00001## ##STR00002## ##STR00003## ##STR00004## ##STR00005## ##STR00006## ##STR00007## ##STR00008## ##STR00009## ##STR00010## ##STR00011## ##STR00012## ##STR00013## ##STR00014## ##STR00015## ##STR00016## ##STR00017## ##STR00018## ##STR00019## ##STR00020## ##STR00021## ##STR00022## ##STR00023## ##STR00024## ##STR00025## ##STR00026## ##STR00027## ##STR00028##

TABLE-US-00021 TABLE 5 Affinities of the trinectin binders to KDR-Fc determined in radioactive equilibrium binding assay Clone KDR (Kd, nM) VR28 11.0 .+-. 0.5 K1 <0.6 .+-. 0.1 K6 <0.9 .+-. 0.1 K9 <1.8 .+-. 0.4 K10 <0.6 .+-. 0.1 K12 <0.6 .+-. 0.1 K13 <0.6 .+-. 0.1 K14 <0.6 .+-. 0.1 K15 <0.4 .+-. 0.1

TABLE-US-00022 TABLE 6 Affinities of the trinectin binders to KDR and Flk-1 determined in radioactive equilibrium binding assay Clone KDR (Kd, nM) Flk-1 (Kd, nM) VR28 11.0 .+-. 0.5 nd* E3 <1.0 .+-. 0.2 5.4 .+-. 1.5 E5 <0.4 .+-. 0.1 3.2 .+-. 0.3 E6 <0.4 .+-. 0.1 7.1 .+-. 1.1 E9 2.4 .+-. 0.3 <1.4 .+-. 0.1 E18 <1.2 .+-. 0.2 <0.5 .+-. 0.1 E19 <1.3 .+-. 0.2 <0.7 .+-. 0.1 E25 <1.6 .+-. 0.4 <1.3 .+-. 0.2 E26 <1.7 .+-. 0.4 2.0 .+-. 0.3 E28 <1.6 .+-. 0.4 2.1 .+-. 0.6 E29 <1.5 .+-. 0.4 <0.9 .+-. 0.2 nd* - binding is not detected at 100 nM of target

TABLE-US-00023 TABLE 7 Determination of ka, kd and Kd by BIAcore assay Clone Target ka (1/M * s) .times. 10.sup.-4 kd(1/s) .times. 10.sup.+5 Kd (nM) E6 KDR 89 6.7 0.08 Flk-1 67 136.0 2.02 E18 KDR 26 12.1 0.46 Flk-1 60 19.5 0.33 E19 KDR 30 1.7 0.06 Flk-1 66 22.3 0.34 E25 KDR 25 5.2 0.21 Flk-1 50 37.8 0.76 E26 KDR 11 5.8 0.51 Flk-1 22 47.7 2.14 E29 KDR 36 7.0 0.19 Flk-1 79 28.8 0.37 M5FL KDR 10 9.2 0.89 Flk-1 28 58.2 2.10 VR28 KDR 3 34 13 159(Q8L) KDR 5 10 2

TABLE-US-00024 TABLE 8 Binding to KDR (CHO KDR) and Flk-1 (CHO Flk-1) expressing cells CHO KDR CHO Flk-1 Clone (EC50, nM) (EC50, nM) E18 4.2 .+-. 1.0 0.9 .+-. 0.4 E19 7.6 .+-. 1.7 5.3 .+-. 2.5 E26 2.6 .+-. 1.2 1.3 .+-. 0.7 E29 2.3 .+-. 1.0 0.6 .+-. 0.1 WT no no

TABLE-US-00025 TABLE 9 Inhibition of VEGF-induced proliferation of KDR (Ba/F3-KDR) and Flk-1 (Ba/F3-Flk) expressing cells Ba/F3-KDR Ba/F3-Flk Clone (IC50, nM) (IC50, nM) E18 5.4 .+-. 1.2 2.4 .+-. 0.2 E19 12.3 .+-. 2.6 5.8 .+-. 1.0 E26 3.2 .+-. 0.5 5.3 .+-. 1.7 E29 10.0 .+-. 2.1 4.7 .+-. 1.2 M5FL 3.9 .+-. 1.1 5.1 .+-. 0.2 WT no no Anti-KDR Ab 17.3 .+-. 7.7 ND Anti-Flk-1 Ab ND 15.0 .+-. 3.2

TABLE-US-00026 TABLE 10 Inhibition of VEGF-induced proliferation of HUVEC cells Clone (IC50, nM) E18 12.8 .+-. 4.6 E19 11.8 .+-. 2.7 E26 14.0 .+-. 5.9 E29 8.4 .+-. 0.8 M5FL 8.5 .+-. 2.8 WT no

TABLE-US-00027 TABLE 11 hKDR Flk-1 k.sub.a k.sub.d k.sub.a k.sub.d (1/Ms) .times. (1/s) .times. K.sub.D (1/Ms) .times. (1/s) .times. K.sub.D 10.sup.-4 10.sup.5 (nM) 10.sup.-4 10.sup.5 (nM) M5FL 7.4 6.7 0.9 14.6 30 2.1 C100 M5FL 20K 0.9 5.4 5.9 2.4 55 22.8 PEG M5FL 40K 0.5 5.9 1.3 1.0 54 57.1 PEG

[0261] All references cited herein are hereby incorporated by reference in their entirety

Sequence CWU 1

1

528111PRTArtificial Sequencebinding polypeptide 1Xaa Gly Xaa Asn Xaa Xaa Glu Leu Xaa Thr Pro 1 5 10211PRTArtificial Sequencebinding polypeptide 2Xaa Glu Arg Asn Gly Arg Xaa Leu Xaa Thr Pro 1 5 10310PRTArtificial Sequencebinding polypeptide 3Xaa Gly Xaa Asn Xaa Arg Xaa Xaa Ile Pro 1 5 10410PRTArtificial Sequencebinding polypeptide 4Xaa Gly Xaa Asn Xaa Arg Xaa Xaa Ile Pro 1 5 10594PRTHomo sapiens 5Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Asp Ala Pro Ala Val Thr Val Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Gly Ser Lys Ser Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Gly Arg Gly Asp65 70 75 80Ser Pro Ala Ser Ser Lys Pro Ile Ser Ile Asn Tyr Arg Thr 85 90686PRTArtificial Sequencebinding polypeptide 6Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Gly Leu Tyr Gly His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 85786PRTArtificial Sequencebinding polypeptide 7Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Glu Asn Gly Gln Phe Leu Leu Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 85886PRTArtificial Sequencebinding polypeptide 8Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Gly Pro Asn Asp Asn Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 85986PRTArtificial Sequencebinding polypeptide 9Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ala Gly Trp Asp Asp His Glu Leu Phe Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851086PRTArtificial Sequencebinding polypeptide 10Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ser Gly His Asn Asp His Met Leu Met Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851186PRTArtificial Sequencebinding polypeptide 11Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ala Gly Tyr Asn Asp Gln Ile Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851286PRTArtificial Sequencebinding polypeptide 12Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Gly Leu Tyr Gly Lys Glu Leu Leu Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851386PRTArtificial Sequencebinding polypeptide 13Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Thr Gly Pro Asn Asp Arg Leu Leu Phe Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851486PRTArtificial Sequencebinding polypeptide 14Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Val Tyr Asn Asp His Glu Ile Lys Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851586PRTArtificial Sequencebinding polypeptide 15Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Lys Asp Gly Arg Val Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851686PRTArtificial Sequencebinding polypeptide 16Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Val His His Asp Arg Glu Ile Lys Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851786PRTArtificial Sequencebinding polypeptide 17Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Gln Ala Pro Asn Asp Arg Val Leu Tyr Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851886PRTArtificial Sequencebinding polypeptide 18Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Arg Glu Glu Asn Asp His Glu Leu Leu Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 851986PRTArtificial Sequencebinding polypeptide 19Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Thr His Asn Gly His Pro Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 852086PRTArtificial Sequencebinding polypeptide 20Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Leu Lys Gly His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 852194PRTArtificial Sequencebinding polypeptide 21Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Ala Gln Asn65 70 75 80Asp His Glu Leu Ile Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 902295PRTArtificial Sequencebinding polypeptide 22Val Ser Asp Val Pro Arg Asp Leu Gln Glu Val Val Ala Ala Thr Pro 1 5 10 15Thr Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr 20 25 30Tyr Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu 35 40 45Phe Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys 50 55 60Pro Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Met Ala Gln65 70 75 80Ser Gly His Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 90 952386PRTArtificial Sequencebinding polypeptide 23Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Lys Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 852486PRTArtificial Sequencebinding polypeptide 24Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 852586PRTArtificial Sequencebinding polypeptide 25Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg His Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 852686PRTArtificial Sequencebinding polypeptide 26Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Met Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 852786PRTArtificial Sequencebinding polypeptide 27Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Arg Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 852886PRTArtificial Sequencebinding polypeptide 28Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Arg Val Leu Asn Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 852986PRTArtificial Sequencebinding polypeptide 29Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Gln Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 853086PRTArtificial Sequencebinding polypeptide 30Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Thr Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg

Thr 853186PRTArtificial Sequencebinding polypeptide 31Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 853286PRTArtificial Sequencebinding polypeptide 32Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Arg Leu Leu Asn Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 853386PRTArtificial Sequencebinding polypeptide 33Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr His Glu Arg Asn Gly Arg Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 853486PRTArtificial Sequencebinding polypeptide 34Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Glu Arg Asn Gly Arg Val Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 853586PRTArtificial Sequencebinding polypeptide 35Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Gln Leu Tyr Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 853686PRTArtificial Sequencebinding polypeptide 36Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Ala Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 853786PRTArtificial Sequencebinding polypeptide 37Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Asn Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 853886PRTArtificial Sequencebinding polypeptide 38Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Arg Val Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 853986PRTArtificial Sequencebinding polypeptide 39Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Val Leu Asn Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854086PRTArtificial Sequencebinding polypeptide 40Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Lys Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854186PRTArtificial Sequencebinding polypeptide 41Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Thr Leu Met Met Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854286PRTArtificial Sequencebinding polypeptide 42Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Glu Arg Asn Gly Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854386PRTArtificial Sequencebinding polypeptide 43Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Glu Arg Asn Gly Arg Thr Leu Arg Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854486PRTArtificial Sequencebinding polypeptide 44Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Lys Thr Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854586PRTArtificial Sequencebinding polypeptide 45Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Asp Arg Val Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854686PRTArtificial Sequencebinding polypeptide 46Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Arg Lys Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854786PRTArtificial Sequencebinding polypeptide 47Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Pro Asn Gly Arg Val Leu Asn Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854886PRTArtificial Sequencebinding polypeptide 48Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Asp Arg Val Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 854986PRTArtificial Sequencebinding polypeptide 49Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Glu Leu Lys Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 855086PRTArtificial Sequencebinding polypeptide 50Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Glu Leu Arg Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 855186PRTArtificial Sequencebinding polypeptide 51Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Gln Glu Arg Asn Gly Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 855286PRTArtificial Sequencebinding polypeptide 52Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 855386PRTArtificial Sequencebinding polypeptide 53Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Val Leu Ser Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 855486PRTArtificial Sequencebinding polypeptide 54Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asp Gly Arg Thr Leu Arg Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 855586PRTArtificial Sequencebinding polypeptide 55Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Glu Leu Asn Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 855686PRTArtificial Sequencebinding polypeptide 56Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Val Leu Ile Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 855786PRTArtificial Sequencebinding polypeptide 57Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Arg Glu Leu Met Val Pro Ile65

70 75 80Ser Ile Asn Tyr Arg Thr 855886PRTArtificial Sequencebinding polypeptide 58Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Arg Asn Asp Arg Lys Leu Met Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 855986PRTArtificial Sequencebinding polypeptide 59Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu His Asn Gly Arg Thr Ser Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856086PRTArtificial Sequencebinding polypeptide 60Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asp Gly Arg Lys Leu Tyr Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856186PRTArtificial Sequencebinding polypeptide 61Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Arg Glu Leu Asn Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856286PRTArtificial Sequencebinding polypeptide 62Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Trp Asn Gly Arg Leu Leu Ser Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856386PRTArtificial Sequencebinding polypeptide 63Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Gln Asn Gly Arg Leu Leu Asn Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856486PRTArtificial Sequencebinding polypeptide 64Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ile Glu Lys Asn Gly Arg His Leu Asn Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856586PRTArtificial Sequencebinding polypeptide 65Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Trp Asn Gly Lys Met Leu Ser Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856686PRTArtificial Sequencebinding polypeptide 66Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Tyr Asn Asp Arg Leu Leu Phe Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856786PRTArtificial Sequencebinding polypeptide 67Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Pro Asn Asp Arg Leu Leu Asn Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856886PRTArtificial Sequencebinding polypeptide 68Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Pro Asn Asn Arg Glu Leu Ile Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 856986PRTArtificial Sequencebinding polypeptide 69Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Leu Asn Gly Lys Tyr Leu Phe Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857086PRTArtificial Sequencebinding polypeptide 70Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Gly Trp Asn Asp Arg Glu Leu Phe Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857186PRTArtificial Sequencebinding polypeptide 71Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Gly Trp Asn Gly Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857286PRTArtificial Sequencebinding polypeptide 72Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Gly Trp Asn Asp Arg Glu Leu Leu Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857386PRTArtificial Sequencebinding polypeptide 73Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Trp Asn Asn Arg Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857486PRTArtificial Sequencebinding polypeptide 74Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Trp Asn Gly Arg Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857586PRTArtificial Sequencebinding polypeptide 75Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asn Glu Arg Asn Gly Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857686PRTArtificial Sequencebinding polypeptide 76Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Lys Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857786PRTArtificial Sequencebinding polypeptide 77Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857886PRTArtificial Sequencebinding polypeptide 78Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Glu Leu Lys Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 857986PRTArtificial Sequencebinding polypeptide 79Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Gln Glu Arg Asn Gly Arg Glu Leu Arg Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858086PRTArtificial Sequencebinding polypeptide 80Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Glu Leu Leu Trp Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858186PRTArtificial Sequencebinding polypeptide 81Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Arg Glu Leu Met Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858286PRTArtificial Sequencebinding polypeptide 82Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Leu Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858386PRTArtificial Sequencebinding polypeptide 83Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Arg Val Leu Ile Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858486PRTArtificial Sequencebinding polypeptide 84Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55

60Tyr Ala Val Thr Val Glu Arg Asn Gly His Lys Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858586PRTArtificial Sequencebinding polypeptide 85Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Glu Arg Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858686PRTArtificial Sequencebinding polypeptide 86Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Gly Pro Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858786PRTArtificial Sequencebinding polypeptide 87Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Gly Pro Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858886PRTArtificial Sequencebinding polypeptide 88Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Gly Lys Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 858987PRTArtificial Sequencebinding polypeptide 89Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15His Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr 20 25 30Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro 35 40 45Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr 50 55 60Gly Tyr Ala Val Thr Val His Trp Asn Gly Arg Glu Leu Met Thr Pro65 70 75 80Ile Ser Ile Asn Tyr Arg Thr 859086PRTArtificial Sequencebinding polypeptide 90Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Glu Trp Asn Gly Arg Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 859186PRTArtificial Sequencebinding polypeptide 91Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly His Thr Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 859286PRTArtificial Sequencebinding polypeptide 92Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Glu Asn Gly Arg Gln Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 859386PRTArtificial Sequencebinding polypeptide 93Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Gly Gln Val Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 859486PRTArtificial Sequencebinding polypeptide 94Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asn Gly Gln Val Leu Tyr Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 859586PRTArtificial Sequencebinding polypeptide 95Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Trp Gly Tyr Lys Asp His Glu Leu Leu Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 859686PRTArtificial Sequencebinding polypeptide 96Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Arg Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 859786PRTArtificial Sequencebinding polypeptide 97Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Pro Asn Asp Arg Leu Leu Asn Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 859886PRTArtificial Sequencebinding polypeptide 98Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Ala Arg Asp Gly His Glu Ile Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 859986PRTArtificial Sequencebinding polypeptide 99Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Gln Asn Gly Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510086PRTArtificial Sequencebinding polypeptide 100Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Glu Asn Gly Arg Val Leu Asn Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510186PRTArtificial Sequencebinding polypeptide 101Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Pro Asn Gly Arg Tyr Leu Met Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510286PRTArtificial Sequencebinding polypeptide 102Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Gly Arg Asn Gly Arg Glu Leu Phe Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510386PRTArtificial Sequencebinding polypeptide 103Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ser Gly Arg Asn Asp Arg Glu Leu Leu Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510486PRTArtificial Sequencebinding polypeptide 104Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Arg Asp Gly Arg Glu Leu Asn Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510586PRTArtificial Sequencebinding polypeptide 105Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Gln Asn Gly Arg Val Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510686PRTArtificial Sequencebinding polypeptide 106Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu His Asn Gly Arg Val Leu Asn Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510786PRTArtificial Sequencebinding polypeptide 107Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Pro Asn Gly Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510886PRTArtificial Sequencebinding polypeptide 108Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Gln Asn Gly Arg Val Leu Asn Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8510986PRTArtificial Sequencebinding polypeptide 109Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Gly Arg Asn Gly His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8511086PRTArtificial Sequencebinding polypeptide 110Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Gly Arg Asn Gly Arg Glu Leu Met Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8511186PRTArtificial Sequencebinding polypeptide 111Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro

Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Arg Asn Asn Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8511286PRTArtificial Sequencebinding polypeptide 112Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Glu Arg Ser Gly Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8511394PRTArtificial Sequencebinding polypeptide 113Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Arg Ala Leu Leu65 70 75 80Ser Ile Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9011494PRTArtificial Sequencebinding polypeptide 114Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Phe Ala Arg Lys65 70 75 80Gly Thr Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9011594PRTArtificial Sequencebinding polypeptide 115Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Glu Arg Cys65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9011694PRTArtificial Sequencebinding polypeptide 116Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Arg Glu Arg Asn65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9011794PRTArtificial Sequencebinding polypeptide 117Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Lys Glu Arg Asn65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9011894PRTArtificial Sequencebinding polypeptide 118Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Cys Glu Arg Asn65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9011994PRTArtificial Sequencebinding polypeptide 119Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Glu Arg Thr65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012094PRTArtificial Sequencebinding polypeptide 120Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Glu Arg Thr65 70 75 80Gly Lys Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012194PRTArtificial Sequencebinding polypeptide 121Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ile Glu Arg Thr65 70 75 80Cys Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012294PRTArtificial Sequencebinding polypeptide 122Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Gly Gly Met Ile65 70 75 80Val Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012394PRTArtificial Sequencebinding polypeptide 123Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Phe Gly Arg Ser65 70 75 80Ser Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012494PRTArtificial Sequencebinding polypeptide 124Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Arg His Lys Ser65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012594PRTArtificial Sequencebinding polypeptide 125Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Arg His Arg Asp65 70 75 80Lys Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012694PRTArtificial Sequencebinding polypeptide 126Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Tyr His Arg Gly65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012794PRTArtificial Sequencebinding polypeptide 127Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Arg His Arg Gly65 70 75 80Cys Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012894PRTArtificial Sequencebinding polypeptide 128Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ser His Arg Leu65 70 75 80Arg Lys Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9012994PRTArtificial Sequencebinding polypeptide 129Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Met His Arg Gln65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013094PRTArtificial Sequencebinding polypeptide 130Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Phe His Arg Arg65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013194PRTArtificial Sequencebinding polypeptide 131Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Phe His Arg Arg65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013294PRTArtificial Sequencebinding polypeptide 132Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ser His Arg Arg65 70 75 80Arg Asn Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013394PRTArtificial Sequencebinding polypeptide 133Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu His Arg Arg65 70 75 80Val Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013494PRTArtificial Sequencebinding polypeptide 134Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Arg His Arg Arg65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013594PRTArtificial Sequencebinding polypeptide 135Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp His Arg Ser65 70 75 80Arg Lys Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013694PRTArtificial Sequencebinding polypeptide 136Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Arg His Arg Ser65 70

75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013794PRTArtificial Sequencebinding polypeptide 137Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val His Arg Thr65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013894PRTArtificial Sequencebinding polypeptide 138Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp His Arg Val65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9013994PRTArtificial Sequencebinding polypeptide 139Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp His Arg Val65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014094PRTArtificial Sequencebinding polypeptide 140Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp His Arg Trp65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014194PRTArtificial Sequencebinding polypeptide 141Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Lys Arg Ser65 70 75 80Gly Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014294PRTArtificial Sequencebinding polypeptide 142Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Arg Leu Xaa Asn65 70 75 80Xaa Val Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014394PRTArtificial Sequencebinding polypeptide 143Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Arg Thr Pro65 70 75 80His Ala Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014494PRTArtificial Sequencebinding polypeptide 144Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Ser Pro His65 70 75 80Ser Val Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014594PRTArtificial Sequencebinding polypeptide 145Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Ser Arg Gln65 70 75 80Lys Ala Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014694PRTArtificial Sequencebinding polypeptide 146Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ser Ser Tyr Ser65 70 75 80Lys Leu Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014794PRTArtificial Sequencebinding polypeptide 147Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Thr Asp Arg65 70 75 80Gly Ser Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014894PRTArtificial Sequencebinding polypeptide 148Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Gly Thr Arg Thr65 70 75 80Arg Ser Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9014994PRTArtificial Sequencebinding polypeptide 149Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Pro Val Ala Gly65 70 75 80Cys Ser Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015094PRTArtificial Sequencebinding polypeptide 150Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Trp Gln Thr65 70 75 80Pro Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015194PRTArtificial Sequencebinding polypeptide 151Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Trp Gln Thr65 70 75 80Pro Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015294PRTArtificial Sequencebinding polypeptide 152Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Glu Arg Asn65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015394PRTArtificial Sequencebinding polypeptide 153Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Glu Trp Asn65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015494PRTArtificial Sequencebinding polypeptide 154Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Lys Glu Arg Asn65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015594PRTArtificial Sequencebinding polypeptide 155Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Leu His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Gly Ala Leu Asn65 70 75 80Thr Ser Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015694PRTArtificial Sequencebinding polypeptide 156Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Phe Gly Arg Glu65 70 75 80Arg Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015794PRTArtificial Sequencebinding polypeptide 157Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Leu His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ser Gly Arg Val65 70 75 80Ser Phe Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015894PRTArtificial Sequencebinding polypeptide 158Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Phe His Arg Arg65 70 75 80Arg Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9015994PRTArtificial Sequencebinding polypeptide 159Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Leu His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Ile Arg Met65 70 75 80Asn Thr Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9016094PRTArtificial Sequencebinding polypeptide 160Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Leu His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Cys Leu His Leu65 70 75 80Ile Thr Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9016194PRTArtificial Sequencebinding polypeptide 161Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Leu His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Leu Lys Leu65 70 75 80Thr Leu Glu Leu Phe Thr Pro Ile Ser Ile

Asn Tyr Arg Thr 85 9016294PRTArtificial Sequencebinding polypeptide 162Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Leu His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Leu Lys Leu65 70 75 80Thr Leu Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9016394PRTArtificial Sequencebinding polypeptide 163Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Phe His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Leu Lys Leu65 70 75 80Thr Leu Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9016494PRTArtificial Sequencebinding polypeptide 164Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Leu His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ala Leu Met Ala65 70 75 80Ser Gly Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9016594PRTArtificial Sequencebinding polypeptide 165Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Leu His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ser Met Lys Asn65 70 75 80Arg Leu Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9016694PRTArtificial Sequencebinding polypeptide 166Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Phe His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Arg Cys Leu65 70 75 80Ile Pro Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9016794PRTArtificial Sequencebinding polypeptide 167Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Leu His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Ser Arg Gln65 70 75 80Lys Ala Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9016894PRTArtificial Sequencebinding polypeptide 168Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Ser Arg Thr65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9016994PRTArtificial Sequencebinding polypeptide 169Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Trp Arg Thr65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017094PRTArtificial Sequencebinding polypeptide 170Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Thr Glu Arg Thr65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017194PRTArtificial Sequencebinding polypeptide 171Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Lys Glu Arg Ser65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017294PRTArtificial Sequencebinding polypeptide 172Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Glu Arg Asn65 70 75 80Asp Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017394PRTArtificial Sequencebinding polypeptide 173Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr His Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Glu Arg Asp65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017494PRTArtificial Sequencebinding polypeptide 174Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Gln Gly Arg His65 70 75 80Lys Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017594PRTArtificial Sequencebinding polypeptide 175Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Leu Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Lys Glu Arg Asn65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017694PRTArtificial Sequencebinding polypeptide 176Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ile Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Met Ala Gln Asn65 70 75 80Asp His Glu Leu Ile Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017794PRTArtificial Sequencebinding polypeptide 177Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ile Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Met Ala Gln Asn65 70 75 80Asp His Glu Leu Ile Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017894PRTArtificial Sequencebinding polypeptide 178Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Thr Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Glu Arg Asn65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9017994PRTArtificial Sequencebinding polypeptide 179Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Thr Val Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Glu Arg Asn65 70 75 80Asp Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9018094PRTArtificial Sequencebinding polypeptide 180Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg Pro Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Thr Val Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Glu Arg Asn65 70 75 80Asp Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9018194PRTArtificial Sequencebinding polypeptide 181Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Pro Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Lys Glu Arg Ser65 70 75 80Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9018294PRTArtificial Sequencebinding polypeptide 182Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Pro Gln Pro Pro Ala Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Glu Arg Asn65 70 75 80Asp Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9018394PRTArtificial Sequencebinding polypeptide 183Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Cys His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Ile Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Met Ala Gln Asn65 70 75 80Asp His Glu Leu Ile Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 90184285DNAArtificial Sequencebinding polypeptide 184atgggcgaag ttgttgctgc gacccccacc agcctactga tcagctggcg ccacccgcac 60ttcccgacta gatattacag gatcacttac ggagaaacag gaggaaatag ccctgtccag 120gagttcactg tgcctctgca gccccccaca gctaccatca gcggccttaa acctggagtt 180gattatacca tcactgtgta tgctgtcact gacggccgga acgggcgcct cctgagcatc 240ccaatttcca ttaattaccg cacagaaatt gacaaaccat gccag 285185264DNAArtificial Sequencebinding polypeptide 185atgggcgaag ttgttgctgc gacccccacc agcctactga tcagctggcg ccacccgcac 60ttcccgacta gatattacag gatcacttac ggagaaacag gaggaaatag ccctgtccag 120gagttcactg tgcctctgca gccccccaca gctaccatca gcggccttaa acctggagtt 180gattatacca tcactgtgta tgctgtcact gacggccgga acgggcgcct cctgagcatc 240ccaatttcca ttaattaccg caca 26418695PRTArtificial Sequencebinding polypeptide 186Met Gly Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp 1 5 10 15Arg His Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu 20 25 30Thr Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro 35 40 45Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile 50 55 60Thr Val Tyr Ala Val Thr Asp Gly Arg Asn Gly Arg Leu Leu Ser Ile65 70 75 80Pro Ile Ser Ile Asn Tyr Arg Thr Glu Ile Asp Lys Pro Ser Gln 85 90 9518795PRTArtificial Sequencebinding polypeptide 187Met Gly Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp 1 5 10 15Arg His Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu 20 25 30Thr Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro 35 40 45Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile 50 55 60Thr Val Tyr Ala Val Thr Asp Gly Arg Asn Gly Arg Leu Leu Ser Ile65

70 75 80Pro Ile Ser Ile Asn Tyr Arg Thr Glu Ile Asp Lys Pro Cys Gln 85 90 95188102PRTArtificial Sequencebinding polypeptide 188Met Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro 1 5 10 15Thr Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr 20 25 30Tyr Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu 35 40 45Phe Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys 50 55 60Pro Gly Val Asp Tyr Thr Ile Thr Val Tyr Ala Val Thr Asp Gly Arg65 70 75 80Asn Gly Arg Leu Leu Ser Ile Pro Ile Ser Ile Asn Tyr Arg Thr Glu 85 90 95Ile Asp Lys Pro Ser Gln 10018988PRTArtificial Sequencebinding polypeptide 189Met Gly Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp 1 5 10 15Arg His Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu 20 25 30Thr Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro 35 40 45Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile 50 55 60Thr Val Tyr Ala Val Thr Asp Gly Trp Asn Gly Arg Leu Leu Ser Ile65 70 75 80Pro Ile Ser Ile Asn Tyr Arg Thr 8519088PRTArtificial Sequencebinding polypeptide 190Met Gly Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp 1 5 10 15Arg His Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu 20 25 30Thr Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro 35 40 45Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile 50 55 60Thr Val Tyr Ala Val Thr Glu Gly Pro Asn Glu Arg Ser Leu Phe Ile65 70 75 80Pro Ile Ser Ile Asn Tyr Arg Thr 8519195PRTArtificial Sequencebinding polypeptide 191Met Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro 1 5 10 15Thr Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr 20 25 30Tyr Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu 35 40 45Phe Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys 50 55 60Pro Gly Val Asp Tyr Thr Ile Thr Val Tyr Ala Val Thr Glu Gly Pro65 70 75 80Asn Glu Arg Ser Leu Phe Ile Pro Ile Ser Ile Asn Tyr Arg Thr 85 90 9519286PRTArtificial Sequencebinding polypeptide 192Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Val 50 55 60Tyr Ala Val Thr Asp Gly Arg Asn Gly Arg Leu Leu Ser Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8519394PRTArtificial Sequencebinding polypeptide 193Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Val Tyr Ala Val Thr Asp Gly Arg Asn65 70 75 80Gly Arg Leu Leu Ser Ile Pro Ile Ser Ile Asn Tyr Arg Thr 85 9019494PRTArtificial Sequencebinding polypeptide 194Gly Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg 1 5 10 15His Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr 20 25 30Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro 35 40 45Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr 50 55 60Val Tyr Ala Val Thr Asp Gly Arg Asn Gly Arg Leu Leu Ser Ile Pro65 70 75 80Ile Ser Ile Asn Tyr Arg Thr Glu Ile Asp Lys Pro Cys Gln 85 9019594PRTArtificial Sequencebinding polypeptide 195Gly Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg 1 5 10 15His Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr 20 25 30Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro 35 40 45Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr 50 55 60Val Tyr Ala Val Thr Asp Gly Arg Asn Gly Arg Leu Leu Ser Ile Pro65 70 75 80Ile Ser Ile Asn Tyr Arg Thr Glu Ile Asp Lys Pro Ser Gln 85 9019686PRTArtificial Sequencebinding polypeptide 196Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Val 50 55 60Tyr Ala Val Thr Glu Gly Pro Asn Glu Arg Ser Leu Phe Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8519794PRTArtificial Sequencebinding polypeptide 197Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Val Tyr Ala Val Thr Glu Gly Pro Asn65 70 75 80Glu Arg Ser Leu Phe Ile Pro Ile Ser Ile Asn Tyr Arg Thr 85 9019810PRTArtificial Sequencebinding polypeptide 198Val Ala Gln Asn Asp His Glu Leu Ile Thr 1 5 10199109PRTArtificial Sequencebinding polypeptide 199Met Gly Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr 1 5 10 15Pro Thr Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg 20 25 30Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln 35 40 45Glu Phe Thr Val Pro Leu Gln Pro Pro Leu Ala Thr Ile Ser Gly Leu 50 55 60Lys Pro Gly Val Asp Tyr Thr Ile Thr Val Tyr Ala Val Thr Lys Glu65 70 75 80Arg Asn Gly Arg Glu Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 90 95Glu Ile Asp Lys Pro Cys Gln His His His His His His 100 10520087PRTArtificial Sequencebinding polypeptide 200Gly Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg 1 5 10 15His Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr 20 25 30Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro 35 40 45Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr 50 55 60Val Tyr Ala Val Thr Asp Gly Trp Asn Gly Arg Leu Leu Ser Ile Pro65 70 75 80Ile Ser Ile Asn Tyr Arg Thr 8520166DNAArtificial Sequencebinding polypeptide 201gcgtaatacg actcactata gggacaatta ctatttacaa ttacaatggt ttctgatgtt 60ccgagg 6620278DNAArtificial Sequencebinding polypeptide 202gcgtaatacg actcactata gggacaatta ctatttacaa ttacaatgga agttgttgct 60gcgaccccca ccagccta 7820357DNAArtificial Sequencebinding polypeptide 203tttttttttt tttttttttt aaatagcgga tgccttgtcg tcgtcgtcct tgtagtc 5720469DNAArtificial Sequencebinding polypeptide 204atggtttctg atgttccgag ggacctggaa gttgttgctg cgacccccac cagcctactg 60atcagctgg 6920566DNAArtificial Sequencebinding polypeptide 205aggcacagtg aactcctgga cagggctatt tcctcctgtt tctccgtaag tgatcctgta 60atatct 6620666DNAArtificial Sequencebinding polypeptide 206agatattaca ggatcactta cggagaaaca ggaggaaata gccctgtcca ggagttcact 60gtgcct 6620763DNAArtificial Sequencebinding polypeptide 207agtgacagca tacacagtga tggtataatc aactccaggt ttaaggccgc tgatggtagc 60tgt 6320863DNAArtificial Sequencebinding polypeptide 208acagctacca tcagcggcct taaacctgga gttgattata ccatcactgt gtatgctgtc 60act 6320977DNAArtificial Sequencebinding polypeptide 209tttttttttt ttttttttta aatagcggat gccttgtcgt cgtcgtcctt gtagtctgtt 60cggtaattaa tggaaat 7721058DNAArtificial Sequencebinding polypeptide 210ttttaaatag cggatgcctt gtcgtcgtcg tccttgtagt ctgttcggta attaatgg 5821130DNAArtificial Sequencebinding polypeptide 211gtgtatgctg tcactatttc cattaattac 3021268DNAArtificial Sequencebinding polypeptide 212taatacgact cactataggg acaattacta tttacaattc tatcaataca atggtgtctg 60atgtgccg 6821372DNAArtificial Sequencebinding polypeptide 213ccaggagatc agcagggagg tcggggtggc agccaccact tccaggtcgc gcggcacatc 60agacaccatt gt 7221464DNAArtificial Sequencebinding polypeptide 214acctccctgc tgatctcctg gcgccatccg cattttccga cccgctatta ccgcatcact 60tacg 6421563DNAArtificial Sequencebinding polypeptide 215cacagtgaac tcctggaccg ggctattgcc tcctgtttcg ccgtaagtga tgcggtaata 60gcg 6321661DNAArtificial Sequencebinding polypeptide 216cggtccagga gttcactgtg ccgctgcagc cgccggcggc taccatcagc ggccttaaac 60c 6121761DNAArtificial Sequencebinding polypeptide 217cggtccagga gttcactgtg ccgnnsnnsn nsnnsnnsgc taccatcagc ggccttaaac 60c 6121859DNAArtificial Sequencebinding polypeptide 218agtgacagca tacacagtga tggtataatc aacgccaggt ttaaggccgc tgatggtag 5921987DNAArtificial Sequencebinding polypeptide 219accatcactg tgtatgctgt cactnnsnns nnsnnsnnsn nsgaactgtt taccccaatt 60tccatcaact accgcacaga ctacaag 8722097DNAArtificial Sequencebinding polypeptide 220aaatagcgga tgcgcgtttg ttctgatctt ccttatttat gtgatgatgg tggtgatgct 60tgtcgtcgtc gtccttgtag tctgtgcggt agttgat 9722151DNAArtificial Sequencebinding polypeptide 221tttttttttt tttttttttt aaatagcgga tgcgcgtttg ttctgatctt c 5122221DNAArtificial Sequencebinding polypeptide 222gcgcgtttgt tctgatcttc c 2122373DNAArtificial Sequencebinding polypeptide 223tgcctcctgt ttcgccgtaa gtgatgcggt aatagcgsnn snnsnnsnns nnsnnsnncc 60agctgatcag cag 7322469DNAArtificial Sequencebinding polypeptide 224gatggtagct gtsnnsnnsn nsnnaggcac agtgaactcc tggacagggc tattgcctcc 60tgtttcgcc 6922568DNAArtificial Sequencebinding polypeptide 225gtgcggtaat taatggaaat tggsnnsnns nnsnnsnnsn nsnnsnnsnn snnagtgaca 60gcatacac 6822621DNAArtificial Sequencebinding polypeptide 226cctcctgttt ctccgtaagt g 2122721DNAArtificial Sequencebinding polypeptide 227cacttacgga gaaacaggag g 2122863DNAArtificial Sequencebinding polypeptide 228acagctacca tcagcggcct taaacctggc gttgattata ccatcactgt gtatgctgtc 60act 6322918DNAArtificial Sequencebinding polypeptide 229agtgacagca tacacagt 1823075DNAArtificial Sequencebinding polypeptide 230tttttttttt tttttttttt aaatagcgga tgccttgtcg tcgtcgtcct tgtagtctgt 60gcggtaatta atgga 7523128DNAArtificial Sequencebinding polypeptide 231tagagaattc atggagagca aggtgctg 2823239DNAArtificial Sequencebinding polypeptide 232agggagagcg tcaggatgag ttccaagttc gtcttttcc 3923328DNAArtificial Sequencebinding polypeptide 233tagagaattc atggagagca aggcgctg 2823439DNAArtificial Sequencebinding polypeptide 234agggagagcg tcaggatgag ttccaagttg gtcttttcc 3923532DNAArtificial Sequencebinding polypeptide 235tagagaattc atgatgtcgt cctggataag gt 3223639DNAArtificial Sequencebinding polypeptide 236agggagagcg tcaggatgag atgttcccga ccggtttta 3923739DNAArtificial Sequencebinding polypeptide 237ggaaaagacg aacttggaac tcatcctgac gctctccct 3923839DNAArtificial Sequencebinding polypeptide 238ggaaaagacc aacttggaac tcatcctgac gctctccct 3923931DNAArtificial Sequencebinding polypeptide 239tagactcgag tcaagagcaa gccacatagc t 3124039DNAArtificial Sequencebinding polypeptide 240taaaaccggt cgggaacatc tcatcctgac gctctccct 39241126PRTArtificial Sequencebinding polypeptide 241Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125242126PRTArtificial Sequencebinding polypeptide 242Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Leu Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125243126PRTArtificial Sequencebinding polypeptide 243Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala

Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Leu Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125244126PRTArtificial Sequencebinding polypeptide 244Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Ala Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Ile Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125245126PRTArtificial Sequencebinding polypeptide 245Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125246126PRTArtificial Sequencebinding polypeptide 246Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Ala Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125247126PRTArtificial Sequencebinding polypeptide 247Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125248126PRTArtificial Sequencebinding polypeptide 248Ala Ile Gln Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125249126PRTArtificial Sequencebinding polypeptide 249Ala Ile Gln Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125250126PRTArtificial Sequencebinding polypeptide 250Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125251126PRTArtificial Sequencebinding polypeptide 251Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125252126PRTArtificial Sequencebinding polypeptide 252Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125253126PRTArtificial Sequencebinding polypeptide 253Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125254126PRTArtificial Sequencebinding polypeptide 254Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Thr Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125255126PRTArtificial Sequencebinding polypeptide 255Glu Ile Val Met Thr Gln Ser Pro Asp Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125256126PRTArtificial Sequencebinding polypeptide 256Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15Gly Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125257125PRTArtificial Sequencebinding polypeptide 257Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125258125PRTArtificial Sequencebinding polypeptide 258Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125259125PRTArtificial Sequencebinding polypeptide 259Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125260125PRTArtificial Sequencebinding polypeptide 260Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125261125PRTArtificial Sequencebinding polypeptide 261Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Ile Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125262125PRTArtificial Sequencebinding polypeptide 262Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Asp Arg Ala Thr Leu Ser Cys Arg Thr Ser Gln Ser Val Ser Ser Ser 20

25 30Ser Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Pro Gly Thr Asp Phe Thr Leu Thr Val Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125263121PRTArtificial Sequencebinding polypeptide 263Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Val Ser Pro Gly 1 5 10 15Gly Arg Ala Ser Leu Pro Cys Arg Ala Ser Gln Thr Val Thr Asn Asn 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Ser Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Phe Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Glu Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ile Ile Ser Arg Leu Glu65 70 75 80Ala Glu Asp Phe Ala Val Tyr Tyr Cys Val Ser Leu Lys Gly Arg Asp 85 90 95Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp Gly Ser Asp Tyr 100 105 110Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120264126PRTArtificial Sequencebinding polypeptide 264Glu Ile Gly Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15Lys Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ile Lys 20 25 30Leu Ala Gly Tyr Gln Gln Lys Leu Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Glu Ala Ser Thr Arg Ala Thr Gly Val Pro Thr Arg Phe Asn Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Asn Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Thr Gly Gly Thr Gly Asn Tyr Tyr 85 90 95Gly Trp Phe Asp Pro Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala 100 105 110Tyr Gly Ser Asp Tyr Glu Asp Asp Asp Asp Xaa Ala Ser Ala 115 120 125265123PRTArtificial Sequencebinding polypeptide 265Glu Ile Val Leu Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys 1 5 10 15Lys Lys Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Val Thr Ser 20 25 30Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Ser Leu Leu Ile 35 40 45Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Cys Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Gly Leu Gln Pro65 70 75 80Glu Asp Ala Ala Val Tyr Tyr Cys Ala Gly Asp Phe Gly Gly Gln Leu 85 90 95Pro Tyr Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp Gly Ser 100 105 110Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120266128PRTArtificial Sequencebinding polypeptide 266Gln Pro Val Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val Phe Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Val 35 40 45Met Ile Tyr Glu Asp Thr Glu Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80Gln Ala Glu Asp Glu Ala Val Tyr Tyr Cys Ala Arg Arg Asn Tyr Gly 85 90 95Ser Gly Ser Trp Ile Asp Tyr Trp Gly Gln Gly Thr Lys Leu Thr Val 100 105 110Leu Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125267126PRTArtificial Sequencebinding polypeptide 267Ala Ile Gln Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Thr Lys 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala Ser Thr Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Thr Gly Gly Thr Gly Asn Tyr Tyr 85 90 95Gly Trp Phe Asp Pro Trp Gly Gln Gly Thr Lys Val Arg Ser Lys Gln 100 105 110Met Ala Ala Thr Thr Arg Thr Thr Thr Thr Arg His Pro Leu 115 120 125268127PRTArtificial Sequencebinding polypeptide 268Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45Leu Ile Tyr Gly Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu65 70 75 80Gln Ala Glu Asp Glu Ala Val Tyr Tyr Cys Ala Ser Ala Ile Pro Gly 85 90 95Thr Arg Ile Leu Arg Asp Trp Gly Gln Gly Thr Lys Leu Thr Val Leu 100 105 110Ala Asp Gly Ser Asp Tyr Arg Thr Thr Thr Thr Arg His Pro Leu 115 120 125269123PRTArtificial Sequencebinding polypeptide 269Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Leu Ala Trp Tyr Gln Gln Arg Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Ile Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly 50 55 60Ser Glu Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Glu Phe Arg Gly Tyr Ala 85 90 95Gly Tyr Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp Gly Ser 100 105 110Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120270125PRTArtificial Sequencebinding polypeptide 270Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Arg Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125271125PRTArtificial Sequencebinding polypeptide 271Gly Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125272125PRTArtificial Sequencebinding polypeptide 272Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Ala Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125273126PRTArtificial Sequencebinding polypeptide 273Glu Ile Val Met Thr Gln Ser Pro Gly Ala Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Gly Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Glu 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125274126PRTArtificial Sequencebinding polypeptide 274Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Gly Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125275126PRTArtificial Sequencebinding polypeptide 275Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Leu Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125276126PRTArtificial Sequencebinding polypeptide 276Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125277126PRTArtificial Sequencebinding polypeptide 277Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Ser Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Ile Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125278126PRTArtificial Sequencebinding polypeptide 278Ala Ile Gln Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Gly Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125279126PRTArtificial Sequencebinding polypeptide 279Lys Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125280126PRTArtificial Sequencebinding polypeptide 280Asp Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115

120 125281126PRTArtificial Sequencebinding polypeptide 281Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125282126PRTArtificial Sequencebinding polypeptide 282Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125283126PRTArtificial Sequencebinding polypeptide 283Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125284126PRTArtificial Sequencebinding polypeptide 284Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125285126PRTArtificial Sequencebinding polypeptide 285Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Ser Val Ala Trp Tyr Arg Lys Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Ile Val Ala Gly Asp 85 90 95His Tyr Phe Asp His Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125286125PRTArtificial Sequencebinding polypeptide 286Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125287125PRTArtificial Sequencebinding polypeptide 287Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125288125PRTArtificial Sequencebinding polypeptide 288Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125289125PRTArtificial Sequencebinding polypeptide 289Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Glu Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125290125PRTArtificial Sequencebinding polypeptide 290Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125291125PRTArtificial Sequencebinding polypeptide 291Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125292125PRTArtificial Sequencebinding polypeptide 292Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Xaa Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125293125PRTArtificial Sequencebinding polypeptide 293Glu Ile Val Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Ser Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Ala Lys Leu Thr Tyr Tyr Gly 85 90 95Ser Gly Arg Asn Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125294126PRTArtificial Sequencebinding polypeptide 294Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ile Lys 20 25 30Leu Ala Trp Tyr Gln Gln Lys Leu Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala Ser Thr Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Thr Gly Gly Thr Gly Asn Tyr Tyr 85 90 95Gly Trp Phe Asp Pro Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125295126PRTArtificial Sequencebinding polypeptide 295Ala Ile Gln Met Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Thr Lys 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala Ser Thr Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Thr Gly Gly Thr Gly Asn Tyr Tyr 85 90 95Gly Trp Phe Asp Pro Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala 100 105 110Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125296125PRTArtificial Sequencebinding polypeptide 296Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Thr Tyr 20 25 30Leu Asn Trp Tyr Gln Leu Lys Pro Gly Glu Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Asn Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Ala Gly Ser Phe Ile Ala Arg Gly 85 90 95Pro Leu Asn Tyr Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125297125PRTArtificial Sequencebinding polypeptide 297Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Thr Tyr 20 25 30Leu Asn Trp Tyr Gln Leu Lys Pro Gly Glu Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Asn Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Ala Gly Ser Phe Ile Ala Arg Gly 85 90 95Pro Leu Asn Tyr Trp Gly Gln Gly Thr Lys Val Glu Ile Lys Ala Asp 100 105 110Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125298128PRTArtificial Sequencebinding polypeptide 298Ser Tyr Glu Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asp His Val Phe Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45Met Ile Tyr Glu Asp Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80Gln Asp Glu Asp Glu Ala Val Tyr Tyr Cys Ala Lys Arg Arg Tyr Val 85 90 95Asn Ala Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Lys Leu Thr Val 100 105 110Leu Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125299128PRTArtificial Sequencebinding polypeptide 299Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asp His Val Phe Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45Met Ile Tyr Glu Asp Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60Ser Gly Ser Lys Ser

Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80Gln Ala Glu Asp Glu Ala Val Tyr Tyr Cys Ala Lys Arg Arg Tyr Val 85 90 95Asn Ala Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Lys Leu Thr Val 100 105 110Leu Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125300128PRTArtificial Sequencebinding polypeptide 300Gln Ser Val Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15Ser Val Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val Phe Trp Tyr Gln Arg His Pro Gly Lys Ala Pro Lys Leu 35 40 45Ile Ile Tyr Glu Asp Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Phe Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80His Thr Glu Asp Glu Ala Val Tyr Tyr Cys Ala Lys Arg Arg Tyr Val 85 90 95Asn Ala Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Lys Val Thr Val 100 105 110Leu Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125301128PRTArtificial Sequencebinding polypeptide 301Gln Ser Val Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15Ser Val Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val Phe Trp Tyr Gln Arg His Pro Gly Lys Ala Pro Lys Leu 35 40 45Ile Ile Tyr Glu Asp Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Phe Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80His Thr Glu Asp Glu Ala Val Tyr Tyr Cys Ala Lys Arg Arg Tyr Val 85 90 95Asn Ala Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Lys Val Thr Val 100 105 110Leu Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125302128PRTArtificial Sequencebinding polypeptide 302Gln Ser Val Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15Ser Val Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val Phe Trp Tyr Gln Arg His Pro Gly Lys Ala Pro Lys Leu 35 40 45Ile Ile Tyr Glu Asp Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Phe Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80His Thr Glu Asp Glu Ala Val Tyr Tyr Cys Ala Lys Arg Arg Tyr Val 85 90 95Asn Ala Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Lys Val Thr Val 100 105 110Leu Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125303128PRTArtificial Sequencebinding polypeptide 303Gln Ser Val Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15Ser Val Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val Phe Trp Tyr Gln Arg His Pro Gly Lys Ala Pro Lys Leu 35 40 45Ile Ile Tyr Glu Asp Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Phe Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80His Thr Glu Asp Glu Ala Val Tyr Tyr Cys Ala Lys Arg Arg Tyr Val 85 90 95Asn Ala Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Lys Val Thr Val 100 105 110Leu Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125304128PRTArtificial Sequencebinding polypeptide 304Ser Tyr Glu Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15Ser Val Ala Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val Phe Trp Tyr Gln Arg His Pro Gly Lys Ala Pro Lys Leu 35 40 45Ile Ile Tyr Glu Asp Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Phe Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75 80His Thr Glu Asp Glu Ala Val Tyr Tyr Cys Ala Lys Arg Arg Tyr Val 85 90 95Asn Ala Gly Trp Phe Asp Tyr Trp Gly Gln Gly Thr Lys Val Thr Val 100 105 110Leu Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125305127PRTArtificial Sequencebinding polypeptide 305Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly Ala Ala Pro Lys Leu 35 40 45Leu Ile Tyr Gly Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu65 70 75 80Gln Ala Glu Asp Glu Ala Val Tyr Tyr Cys Ala Ser Ala Ile Pro Gly 85 90 95Thr Arg Ile Leu Arg Asp Trp Gly Gln Gly Thr Lys Leu Thr Val Leu 100 105 110Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120 125306123PRTArtificial Sequencebinding polypeptide 306Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15Glu Arg Ala Ala Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asp Lys 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Ala Thr Glu Phe Arg Gly Tyr Ala 85 90 95Gly Tyr Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp Gly Ser 100 105 110Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120307123PRTArtificial Sequencebinding polypeptide 307Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Gly Ala Ser Arg Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Ala Gly Phe Asp Pro Tyr Gln Pro 85 90 95Pro Arg Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp Gly Ser 100 105 110Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120308123PRTArtificial Sequencebinding polypeptide 308Glu Ile Val Leu Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys 1 5 10 15Glu Lys Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Val Thr Ser 20 25 30Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Ser Leu Leu Ile 35 40 45Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Gly Leu Gln Pro65 70 75 80Glu Asp Ala Ala Val Tyr Tyr Cys Ala Gly Asp Phe Gly Gly Gln Leu 85 90 95Pro Tyr Trp Gly Gln Gly Thr Lys Leu Glu Ile Lys Ala Asp Gly Ser 100 105 110Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser Ala 115 120309129PRTArtificial Sequencebinding polypeptide 309Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly 1 5 10 15Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Tyr Ser 20 25 30Asp Gly Asn Thr Tyr Leu Asn Trp Phe Gln Gln Arg Pro Gly Gln Ser 35 40 45Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Ala Val Tyr Tyr Cys Ala Arg Ala 85 90 95Gly Ser Ser Trp Lys Phe Asp Tyr Trp Gly Gln Gly Thr Lys Val Glu 100 105 110Ile Lys Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser 115 120 125Ala310129PRTArtificial Sequencebinding polypeptide 310Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly 1 5 10 15Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Leu Thr 20 25 30Ser Gly Asp Asn Tyr Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ala Arg Met 85 90 95Gly Ala Val Ala Gly Tyr Asp Tyr Trp Gly Gln Gly Thr Lys Val Glu 100 105 110Ile Lys Ala Asp Gly Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Ser 115 120 125Ala31186PRTArtificial Sequencebinding polypeptide 311Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Gln Asp Gly His Val Leu Tyr Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8531286PRTArtificial Sequencebinding polypeptide 312Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Gly Lys Asn Gly His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8531386PRTArtificial Sequencebinding polypeptide 313Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Pro Gly Pro Gly Asp Arg Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8531486PRTArtificial Sequencebinding polypeptide 314Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ala Gly Pro Gly Ala His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8531586PRTArtificial Sequencebinding polypeptide 315Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Asn Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8531686PRTArtificial Sequencebinding polypeptide 316Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Tyr Gly Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8531786PRTArtificial Sequencebinding polypeptide 317Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala His Asn Gly Asn Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8531886PRTArtificial Sequencebinding polypeptide 318Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Trp Asn Gly His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8531986PRTArtificial Sequencebinding polypeptide 319Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Leu Arg Asp Arg Glu Leu Phe Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532086PRTArtificial Sequencebinding polypeptide 320Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ser Gly Leu Asn Asp Arg Val Leu Phe Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532186PRTArtificial Sequencebinding polypeptide 321Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr

Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Gly Pro Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532286PRTArtificial Sequencebinding polypeptide 322Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly His Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532386PRTArtificial Sequencebinding polypeptide 323Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Leu Asn Asp Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532486PRTArtificial Sequencebinding polypeptide 324Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Gly His Lys Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532586PRTArtificial Sequencebinding polypeptide 325Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val His Trp Asn Gly His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532686PRTArtificial Sequencebinding polypeptide 326Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Gly Phe Met Ala His Glu Leu Met Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532786PRTArtificial Sequencebinding polypeptide 327Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ala Gly Leu Asn Glu His Glu Leu Leu Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532886PRTArtificial Sequencebinding polypeptide 328Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Asp Asn Ala Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8532986PRTArtificial Sequencebinding polypeptide 329Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Lys Asp Val Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533086PRTArtificial Sequencebinding polypeptide 330Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ser Asp Ser Gly His Ala Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533186PRTArtificial Sequencebinding polypeptide 331Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Pro Tyr Glu His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533286PRTArtificial Sequencebinding polypeptide 332Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ala Gly Arg His Asp His Glu Leu Ile Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533386PRTArtificial Sequencebinding polypeptide 333Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ile Gly Pro Asn Asn His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533486PRTArtificial Sequencebinding polypeptide 334Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Gln Asn Gly Arg Glu Leu Ile Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533586PRTArtificial Sequencebinding polypeptide 335Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ala Gly Leu Asp Glu His Glu Leu Leu Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533686PRTArtificial Sequencebinding polypeptide 336Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Pro Asn Gly His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533786PRTArtificial Sequencebinding polypeptide 337Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Gly His Ala Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533886PRTArtificial Sequencebinding polypeptide 338Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Tyr Asn Asn Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8533986PRTArtificial Sequencebinding polypeptide 339Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asp Gly His Phe Leu Tyr Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534086PRTArtificial Sequencebinding polypeptide 340Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ser Gly His Asn Gly His Glu Val Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534186PRTArtificial Sequencebinding polypeptide 341Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Asp Gln Ser Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534286PRTArtificial Sequencebinding polypeptide 342Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Pro Asn Glu Arg Met Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534386PRTArtificial Sequencebinding polypeptide 343Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Gly Tyr Tyr Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534486PRTArtificial Sequencebinding polypeptide 344Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Thr His Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534586PRTArtificial Sequencebinding polypeptide 345Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Arg Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534686PRTArtificial Sequencebinding polypeptide 346Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Trp Ala Gln Asn Gly Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534786PRTArtificial Sequencebinding polypeptide 347Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Lys Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534886PRTArtificial Sequencebinding polypeptide 348Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5

10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Pro Asn Asp His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8534986PRTArtificial Sequencebinding polypeptide 349Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Thr Gly Trp Asn Gly Asn Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535086PRTArtificial Sequencebinding polypeptide 350Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala His Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535186PRTArtificial Sequencebinding polypeptide 351Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Glu Gln Asn Asp Arg Val Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535286PRTArtificial Sequencebinding polypeptide 352Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Thr Gly His His Asp His Glu Leu Ile Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535386PRTArtificial Sequencebinding polypeptide 353Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala His Glu Asn Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535486PRTArtificial Sequencebinding polypeptide 354Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Leu Asn Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535586PRTArtificial Sequencebinding polypeptide 355Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ala Gly Pro Asn Asp His Gln Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535686PRTArtificial Sequencebinding polypeptide 356Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Ala Met Tyr Gly Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535786PRTArtificial Sequencebinding polypeptide 357Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Trp Asp Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535886PRTArtificial Sequencebinding polypeptide 358Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Gly Gln Asn Asp Lys Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8535986PRTArtificial Sequencebinding polypeptide 359Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Gly His Glu Leu Tyr Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536086PRTArtificial Sequencebinding polypeptide 360Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Pro Gly His Asn Asp His Glu Leu Met Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536186PRTArtificial Sequencebinding polypeptide 361Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Ala Arg Asn Gly Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536286PRTArtificial Sequencebinding polypeptide 362Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala His Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536386PRTArtificial Sequencebinding polypeptide 363Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala His Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536486PRTArtificial Sequencebinding polypeptide 364Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Gln Asn Asp Arg Gln Leu Leu Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536586PRTArtificial Sequencebinding polypeptide 365Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ala Gly Gly Asn Gly His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536686PRTArtificial Sequencebinding polypeptide 366Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr His Gly Pro Tyr Asp Gln Val Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536786PRTArtificial Sequencebinding polypeptide 367Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ile Glu Gln Ser Gly Leu Gln Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536886PRTArtificial Sequencebinding polypeptide 368Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8536986PRTArtificial Sequencebinding polypeptide 369Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Trp Asp Gly Arg Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8537086PRTArtificial Sequencebinding polypeptide 370Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Tyr Asn Gly Arg Glu Ile Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8537186PRTArtificial Sequencebinding polypeptide 371Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Trp Ser Gln Asn Asn Arg Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8537286PRTArtificial Sequencebinding polypeptide 372Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Thr Trp Asn Asp His Glu Ile Arg Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8537386PRTArtificial Sequencebinding polypeptide 373Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Gly His Gln Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8537494PRTArtificial Sequencebinding polypeptide 374Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Thr His Asn65 70 75 80Gly His Pro Leu Met Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 90375107PRTArtificial

Sequencebinding polypeptide 375Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Tyr Tyr Arg Ile Thr 35 40 45Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro 50 55 60Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp65 70 75 80Tyr Thr Ile Thr Gly Tyr Ala Val Thr Phe Ala Gln Asn Asp His Gln 85 90 95Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 100 105376107PRTArtificial Sequencebinding polypeptide 376Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Tyr Tyr Arg Ile Thr 35 40 45Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro 50 55 60Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp65 70 75 80Tyr Thr Ile Thr Gly Tyr Ala Val Thr Gly Gly Gln Met Asn Arg Val 85 90 95Leu Met Thr Pro Ile Ser Ile Asn Tyr Arg Thr 100 105377107PRTArtificial Sequencebinding polypeptide 377Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Tyr Tyr Arg Ile Thr 35 40 45Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro 50 55 60Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp65 70 75 80Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Val His Asn Asp Arg Glu 85 90 95Leu Leu Thr Pro Ile Ser Ile Asn Tyr Arg Thr 100 105378107PRTArtificial Sequencebinding polypeptide 378Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Tyr Tyr Arg Ile Thr 35 40 45Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro 50 55 60Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp65 70 75 80Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Ala Gln Asn Gly His Glu 85 90 95Leu Phe Thr Pro Ile Ser Ile Asn Tyr Arg Thr 100 105379115PRTArtificial Sequencebinding polypeptide 379Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser 50 55 60Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile65 70 75 80Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val 85 90 95Thr Val His Trp Asn Gly His Glu Leu Met Thr Pro Ile Ser Ile Asn 100 105 110Tyr Arg Thr 115380107PRTArtificial Sequencebinding polypeptide 380Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Tyr Tyr Arg Ile Thr 35 40 45Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro 50 55 60Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp65 70 75 80Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Gly Trp Asn Asp His Glu 85 90 95Leu Tyr Ile Pro Ile Ser Ile Asn Tyr Arg Thr 100 10538186PRTArtificial Sequencebinding polypeptide 381Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Ala Gly His Lys Asp Gln Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8538286PRTArtificial Sequencebinding polypeptide 382Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asn His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8538386PRTArtificial Sequencebinding polypeptide 383Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Trp Asn Asp His Glu Ile Tyr Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8538486PRTArtificial Sequencebinding polypeptide 384Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Thr Gly Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8538586PRTArtificial Sequencebinding polypeptide 385Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Trp Ser Gly His Glu Leu Phe Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8538686PRTArtificial Sequencebinding polypeptide 386Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly His Asn Asp Arg Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8538786PRTArtificial Sequencebinding polypeptide 387Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Trp Asn Gln Asn Gly Gln Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8538886PRTArtificial Sequencebinding polypeptide 388Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Gly Gln Asn Gly His Ala Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8538986PRTArtificial Sequencebinding polypeptide 389Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Arg Gly Leu Asn Asp Gly Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539086PRTArtificial Sequencebinding polypeptide 390Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Gly Pro Ser Asp His Val Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539186PRTArtificial Sequencebinding polypeptide 391Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asn His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539286PRTArtificial Sequencebinding polypeptide 392Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp His Lys Leu Phe Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539386PRTArtificial Sequencebinding polypeptide 393Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Arg Asp Gln Tyr Glu His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539486PRTArtificial Sequencebinding polypeptide 394Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Leu Asn Gly His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539586PRTArtificial Sequencebinding polypeptide 395Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Glu Ser Asn Gly His Ala Leu Phe Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539686PRTArtificial Sequencebinding polypeptide 396Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Asn Asn His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539786PRTArtificial Sequencebinding polypeptide 397Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Trp Asp Gln Asn Gly His Val Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539886PRTArtificial Sequencebinding polypeptide 398Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Gly Leu Asn Asp His Glu Leu Ile Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8539986PRTArtificial Sequencebinding polypeptide 399Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Gly Leu Asn Asp His Glu Leu Met Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8540086PRTArtificial Sequencebinding polypeptide 400Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25

30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Gly Gln Asn Asp Gln Leu Leu Phe Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8540186PRTArtificial Sequencebinding polypeptide 401Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Gly His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8540286PRTArtificial Sequencebinding polypeptide 402Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8540386PRTArtificial Sequencebinding polypeptide 403Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Gly His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8540486PRTArtificial Sequencebinding polypeptide 404Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Glu Arg Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8540586PRTArtificial Sequencebinding polypeptide 405Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Trp Asn Asp His Met Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8540686PRTArtificial Sequencebinding polypeptide 406Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Pro Asn Asp Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8540794PRTArtificial Sequencebinding polypeptide 407Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Gly Pro Asn65 70 75 80Glu Arg Met Leu Met Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9040886PRTArtificial Sequencebinding polypeptide 408Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala His Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8540986PRTArtificial Sequencebinding polypeptide 409Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Lys Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541086PRTArtificial Sequencebinding polypeptide 410Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Trp Ala Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541186PRTArtificial Sequencebinding polypeptide 411Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Ala Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541286PRTArtificial Sequencebinding polypeptide 412Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Glu Gln Asn Gly His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541386PRTArtificial Sequencebinding polypeptide 413Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Asp Ala Pro Asn Gly Arg Glu Leu Met Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541486PRTArtificial Sequencebinding polypeptide 414Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Gly Gly Arg Asn Gly His Thr Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541586PRTArtificial Sequencebinding polypeptide 415Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ser Gln Thr Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541686PRTArtificial Sequencebinding polypeptide 416Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Asn Glu His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541786PRTArtificial Sequencebinding polypeptide 417Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Gly His Glu Leu Lys Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541886PRTArtificial Sequencebinding polypeptide 418Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp Arg Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8541986PRTArtificial Sequencebinding polypeptide 419Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Asn His His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8542086PRTArtificial Sequencebinding polypeptide 420Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Pro His Asp Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8542194PRTArtificial Sequencebinding polypeptide 421Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Met Gly Phe Met65 70 75 80Ala His Glu Leu Met Val Pro Ile Ser Ile Asn Tyr Arg Thr 85 9042286PRTArtificial Sequencebinding polypeptide 422Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8542386PRTArtificial Sequencebinding polypeptide 423Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Val Arg Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8542486PRTArtificial Sequencebinding polypeptide 424Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asp Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8542586PRTArtificial Sequencebinding polypeptide 425Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Asp Ile Arg Val Leu Trp Leu Asn Thr Thr Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8542686PRTArtificial Sequencebinding polypeptide 426Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Thr Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8542786PRTArtificial Sequencebinding polypeptide 427Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1

5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8542886PRTArtificial Sequencebinding polypeptide 428Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Asp His Lys Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8542986PRTArtificial Sequencebinding polypeptide 429Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543086PRTArtificial Sequencebinding polypeptide 430Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543186PRTArtificial Sequencebinding polypeptide 431Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asn His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543286PRTArtificial Sequencebinding polypeptide 432Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asp Arg Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543386PRTArtificial Sequencebinding polypeptide 433Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Asn Asn His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543486PRTArtificial Sequencebinding polypeptide 434Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Gly His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543586PRTArtificial Sequencebinding polypeptide 435Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Thr Ala His Asn Gly His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543686PRTArtificial Sequencebinding polypeptide 436Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Tyr His Asp His Ala Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543786PRTArtificial Sequencebinding polypeptide 437Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Trp Ala Trp Asn Asp His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543886PRTArtificial Sequencebinding polypeptide 438Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8543986PRTArtificial Sequencebinding polypeptide 439Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Asp His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8544086PRTArtificial Sequencebinding polypeptide 440Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8544186PRTArtificial Sequencebinding polypeptide 441Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Gly His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8544294PRTArtificial Sequencebinding polypeptide 442Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Glu Gly Trp Ile65 70 75 80Asp His Glu Ile Met Ile Pro Ile Ser Ile Asn Tyr Arg Thr 85 9044386PRTArtificial Sequencebinding polypeptide 443Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Gly Gln Asn Gly Ser Glu Leu Ile Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8544486PRTArtificial Sequencebinding polypeptide 444Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Asp Arg Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8544586PRTArtificial Sequencebinding polypeptide 445Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Asn Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8544686PRTArtificial Sequencebinding polypeptide 446Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Ser Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8544786PRTArtificial Sequencebinding polypeptide 447Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Asp Arg Asn Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8544886PRTArtificial Sequencebinding polypeptide 448Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asp His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8544986PRTArtificial Sequencebinding polypeptide 449Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8545086PRTArtificial Sequencebinding polypeptide 450Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Glu Asn Asp Arg Lys Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8545186PRTArtificial Sequencebinding polypeptide 451Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8545286PRTArtificial Sequencebinding polypeptide 452Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Gly Pro Asn Gly His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8545386PRTArtificial Sequencebinding polypeptide 453Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Val Arg Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr

8545486PRTArtificial Sequencebinding polypeptide 454Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Thr Gln Asn Gly His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8545586PRTArtificial Sequencebinding polypeptide 455Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Thr Gln Asn Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8545686PRTArtificial Sequencebinding polypeptide 456Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Gly His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8545786PRTArtificial Sequencebinding polypeptide 457Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp Arg Gln Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8545886PRTArtificial Sequencebinding polypeptide 458Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Pro Asn Asp Arg Glu Leu Ile Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8545986PRTArtificial Sequencebinding polypeptide 459Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Glu His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8546086PRTArtificial Sequencebinding polypeptide 460Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asn His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8546186PRTArtificial Sequencebinding polypeptide 461Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Glu Ala His His Gly His Glu Leu Met Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8546286PRTArtificial Sequencebinding polypeptide 462Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Gly Asp His Asn Asp Arg Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8546394PRTArtificial Sequencebinding polypeptide 463Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Gly Gly Gln Met65 70 75 80Asn Arg Val Leu Met Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9046486PRTArtificial Sequencebinding polypeptide 464Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala His Asn Asp Arg Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8546586PRTArtificial Sequencebinding polypeptide 465Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Pro Gln Asn Gly His Glu Leu Ile Thr Met Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8546686PRTArtificial Sequencebinding polypeptide 466Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8546786PRTArtificial Sequencebinding polypeptide 467Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Asp Gln Asn Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8546894PRTArtificial Sequencebinding polypeptide 468Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Ala Trp Asn65 70 75 80Asp His Met Leu Met Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9046994PRTArtificial Sequencebinding polypeptide 469Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ser Gly His Asn65 70 75 80Asp His Met Leu Met Ile Pro Ile Ser Ile Asn Tyr Arg Thr 85 9047086PRTArtificial Sequencebinding polypeptide 470Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Gly His Val Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8547186PRTArtificial Sequencebinding polypeptide 471Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Thr His Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8547286PRTArtificial Sequencebinding polypeptide 472Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Asn Asp His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8547386PRTArtificial Sequencebinding polypeptide 473Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asp His Glu Ile Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8547486PRTArtificial Sequencebinding polypeptide 474Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8547586PRTArtificial Sequencebinding polypeptide 475Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ser Gln Gln Asn His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8547686PRTArtificial Sequencebinding polypeptide 476Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8547786PRTArtificial Sequencebinding polypeptide 477Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Tyr Asn Glu His Glu Leu Tyr Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8547886PRTArtificial Sequencebinding polypeptide 478Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln His Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8547986PRTArtificial Sequencebinding polypeptide 479Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Asn Asp Gln Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8548086PRTArtificial Sequencebinding polypeptide 480Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile

Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Arg Asn Asp His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8548186PRTArtificial Sequencebinding polypeptide 481Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Pro Thr Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8548286PRTArtificial Sequencebinding polypeptide 482Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Leu Thr Asp His Val Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8548386PRTArtificial Sequencebinding polypeptide 483Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asp Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8548486PRTArtificial Sequencebinding polypeptide 484Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Ala Gln Asn Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8548586PRTArtificial Sequencebinding polypeptide 485Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Trp Asn Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8548686PRTArtificial Sequencebinding polypeptide 486Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8548786PRTArtificial Sequencebinding polypeptide 487Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Gln Glu Asn Gln Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8548894PRTArtificial Sequencebinding polypeptide 488Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Ala Pro Ser65 70 75 80Ala Arg Glu Leu Met Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9048986PRTArtificial Sequencebinding polypeptide 489Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Val His Asn Gly His Glu Ile Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8549086PRTArtificial Sequencebinding polypeptide 490Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Gly Tyr Glu Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8549194PRTArtificial Sequencebinding polypeptide 491Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Glu Gly Tyr Gln65 70 75 80Asn His Glu Leu Ser Val Pro Ile Ser Ile Asn Tyr Arg Thr 85 9049286PRTArtificial Sequencebinding polypeptide 492Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Asp Gln Asn Asp His Glu Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8549386PRTArtificial Sequencebinding polypeptide 493Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Ser Asp His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8549486PRTArtificial Sequencebinding polypeptide 494Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Asn Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8549586PRTArtificial Sequencebinding polypeptide 495Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8549686PRTArtificial Sequencebinding polypeptide 496Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Gly His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8549786PRTArtificial Sequencebinding polypeptide 497Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Arg Ala Gln Asn Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8549886PRTArtificial Sequencebinding polypeptide 498Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Ser Asn His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8549986PRTArtificial Sequencebinding polypeptide 499Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp Arg Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8550086PRTArtificial Sequencebinding polypeptide 500Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Thr His Asn Glu Gln Tyr Leu Phe Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8550194PRTArtificial Sequencebinding polypeptide 501Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Glu Ile Tyr Asn65 70 75 80Asp His Glu Leu Met Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9050286PRTArtificial Sequencebinding polypeptide 502Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8550386PRTArtificial Sequencebinding polypeptide 503Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ser Gln Tyr Gly His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8550486PRTArtificial Sequencebinding polypeptide 504Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Lys Asn Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8550586PRTArtificial Sequencebinding polypeptide 505Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asn His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8550686PRTArtificial Sequencebinding polypeptide 506Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln His Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8550794PRTArtificial Sequencebinding polypeptide 507Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr

20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Ser His Tyr65 70 75 80Gly Lys Glu Leu Arg Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9050886PRTArtificial Sequencebinding polypeptide 508Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Ala His Glu Leu Met Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8550986PRTArtificial Sequencebinding polypeptide 509Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Gly Gln Asn Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8551086PRTArtificial Sequencebinding polypeptide 510Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asn Asp His Glu Leu Lys Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8551186PRTArtificial Sequencebinding polypeptide 511Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Gly Glu Gln Asn Asp Tyr Glu Leu Leu Val Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8551286PRTArtificial Sequencebinding polypeptide 512Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Leu Thr Gln His Asp His Glu Leu Leu Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8551386PRTArtificial Sequencebinding polypeptide 513Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Met Ala Gln Asn Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8551494PRTArtificial Sequencebinding polypeptide 514Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Glu Ala Pro Asn65 70 75 80Gly Arg Glu Leu Arg Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9051594PRTArtificial Sequencebinding polypeptide 515Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Gly Pro Thr65 70 75 80Asp His Glu Leu Leu Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9051686PRTArtificial Sequencebinding polypeptide 516Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Gly Gln Tyr Asp His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8551786PRTArtificial Sequencebinding polypeptide 517Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Val Ala Gln Asp Glu His Glu Leu Ile Thr Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8551894PRTArtificial Sequencebinding polypeptide 518Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Asp Ala Gln Asn65 70 75 80Val Gln Ala Pro Ile Ala Gln Ile Ser Ile Asn Tyr Arg Thr 85 9051994PRTArtificial Sequencebinding polypeptide 519Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ser Gly Gln Asn65 70 75 80Asp His Ala Leu Leu Ile Pro Ile Ser Ile Asn Tyr Arg Thr 85 9052094PRTArtificial Sequencebinding polypeptide 520Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Glu Asp Ile Arg65 70 75 80Val Leu Trp Leu Asn Thr Thr Ile Ser Ile Asn Tyr Arg Thr 85 9052194PRTArtificial Sequencebinding polypeptide 521Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Trp Asp Gln Asn65 70 75 80Gly His Val Leu Leu Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9052294PRTArtificial Sequencebinding polypeptide 522Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Gly Leu Arg65 70 75 80Asp Arg Glu Leu Phe Val Pro Ile Ser Ile Asn Tyr Arg Thr 85 9052394PRTArtificial Sequencebinding polypeptide 523Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Val Glu Pro Asn65 70 75 80Gly His Lys Leu Ala Ile Pro Ile Ser Ile Asn Tyr Arg Thr 85 9052486PRTArtificial Sequencebinding polypeptide 524Glu Val Val Ala Ala Thr Pro Thr Ser Leu Leu Ile Ser Trp Arg His 1 5 10 15Pro His Phe Pro Thr Arg Tyr Tyr Arg Ile Thr Tyr Gly Glu Thr Gly 20 25 30Gly Asn Ser Pro Val Gln Glu Phe Thr Val Pro Leu Gln Pro Pro Thr 35 40 45Ala Thr Ile Ser Gly Leu Lys Pro Gly Val Asp Tyr Thr Ile Thr Gly 50 55 60Tyr Ala Val Thr Phe Gly Gln Asn Gly Lys Glu Phe Arg Ile Pro Ile65 70 75 80Ser Ile Asn Tyr Arg Thr 8552594PRTArtificial Sequencebinding polypeptide 525Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Ser Gly His Asn65 70 75 80Gly His Glu Val Met Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9052694PRTArtificial Sequencebinding polypeptide 526Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Gly Trp Asn65 70 75 80Asp His Glu Leu Tyr Ile Pro Ile Ser Ile Asn Tyr Arg Thr 85 9052794PRTArtificial Sequencebinding polypeptide 527Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Gly Lys Asp65 70 75 80Val Arg Glu Leu Leu Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 9052894PRTArtificial Sequencebinding polypeptide 528Val Ser Asp Val Pro Arg Asp Leu Glu Val Val Ala Ala Thr Pro Thr 1 5 10 15Ser Leu Leu Ile Ser Trp Arg His Pro His Phe Pro Thr Arg Tyr Tyr 20 25 30Arg Ile Thr Tyr Gly Glu Thr Gly Gly Asn Ser Pro Val Gln Glu Phe 35 40 45Thr Val Pro Leu Gln Pro Pro Thr Ala Thr Ile Ser Gly Leu Lys Pro 50 55 60Gly Val Asp Tyr Thr Ile Thr Gly Tyr Ala Val Thr Leu Ala Leu Phe65 70 75 80Asp His Glu Leu Leu Thr Pro Ile Ser Ile Asn Tyr Arg Thr 85 90

Claims

1. A sustained-release intraocular drug delivery system comprising: a therapeutic component comprising an antiangiogenic polypeptide component; and a polymeric component associated with the therapeutic component to permit the therapeutic component to be released into the interior of an eye of an individual at a therapeutically effective dosage for a period of time after the drug delivery system is placed in the eye.

2. The system of claim 1 wherein said therapeutic component and said polymeric component are combined in a form selected from the group consisting of a) an implant device, or b) a plurality of particles.

3. The system of claim 2 wherein the antiangiogenic polypeptide component comprises an antibody, antibody fragment, or artificial antibody, and humanized versions of these polypeptides.

4. The system of claim 3 wherein the antiangiogenic component comprises an artificial antibody or a humanized version thereof.

5. The system of claim 4 wherein the artificial antibody comprises a scaffold region based upon a fibronectin.

6. The system of claim 5 wherein the artificial antibody comprises fibronectin based "addressable" therapeutic binding molecule ("FATBIM").

7. The system of claim 6 wherein the FATBIM is selected from the group consisting of CT322, C7S100 and C7C100.

8. A sustained-release intraocular drug delivery system comprising: a therapeutic component comprising an antiangiogenic polypeptide component, wherein the therapeutic component is selected from the group consisting of C7S100 and C7C100; and a polymeric component associated with the therapeutic component to permit the therapeutic component to be released into the interior of an eye of an individual at a therapeutically effective dosage for a period of time after the drug delivery system is placed in the eye.

9. A method of treating a retinopathy, the method comprising administering, to a patient in need thereof, a therapeutically effective amount of a polypeptide that binds to human VEGFR-2, the polypeptide comprising between about 80 and about 150 amino acids that have a structural organization comprising: i) at least five to seven beta strands or beta-like strands distributed among at least two beta sheets, and ii) at least one loop portion connecting two strands that are beta strands or beta-like strands, which loop portion participates in binding to VEGFR-2, wherein the polypeptide binds to an extracellular domain of the human VEGFR-2 protein with a dissociation constant (K.sub.D) of less than 1.times.10.sup.-6 M and inhibits VEGFR-2 mediated angiogenesis.